JPH04264254A - Automatic control system for improved type electrophoresis - Google Patents

Abstract

PURPOSE: To improve cataphoresis separation by receiving the support of a computer and utilizing a knowledge based on calibration data that are obtained by separating a gigantic molecule with a known size under conditions for optimizing separation. CONSTITUTION: A system 100 consisting of a computer system 150, a contour fixing homogeneous electric field(CHEF) mapper 120, and a cataphoresis device 130 operates in a complete automatic mode. A user inputs the minimum and maximum fragment sizes of a gigantic molecule to be separated, especially DNA. Related knowledge base is provided, thus determining optimized parameters of a forward voltage gradient, a backward voltage gradient, a voltage gradient, an angle, an initial switch time, a final switch time, and a run period. The mapper 120 can directly calibrate the device 130 using the parameters, thus separating the maximum gigantic molecule.

Description

DETAILED DESCRIPTION OF THE INVENTION [Field of Industrial Application] The present invention generally relates to gel electrophoresis,
More specifically, contour-fixed homogeneous field electrophoresis (conto
ur-clamped homogeneous el
electric field electrophore
sis). [Prior Art] DNA and other macromolecules are separated, fixed, and
The standard method used to read and purify agarose
By electrophoresis through agarose gel
be. This technique is not well separated by other purification techniques.
Effectively and quickly separates a mixture of DNA fragments that cannot be separated.
I can understand it. The gel containing the DNA sample is subjected to an electric field
. For this reason, individual macromolecules within the DNA are transported through the gel.
Moving. different DNA fragments move at different speeds
The visible “bands” of DNA are
occurs within the file. The position of individual DNA fragments within the gel is
can be read from the band. [0003] Specific characteristics of DNA include its size and charge.
(charage), and conformation (conformat)
ion), which affects the speed of movement, and for this reason
affects the separation that is achieved. For example, linear
) and duplex DNA macromolecules are typical
at a rate inversely proportional to the log10 of their molecular weight
transfer through the gel matrix.
move. Nevertheless, closed circular
circular), nicked c
ircular), and linear type (linear f
orms) of the same molecular weight but different conformations
DNA macromolecules pass through an agarose gel at different speeds.
and move. In addition to DNA properties, electrophoretic parameters
It is also known as macromolecular migration (migration).
on) (mobility).
Ru. DNA fragments of a given size are separated from each other in agarose.
move at different speeds through a gel containing different concentrations. dark
By changing the degree of
It is possible to decompose it into different ranges. closed circular form of DNA
applied field strength (voltage gradient), buffer
The ionic strength of the buffer and the
super helical twist
The density of st) also affects migration. low electricity
At pressure, the mobility of linear DNA fragments (cm/hr) is
proportional to the voltage available. However, this linear relationship
collapses at higher potentials for polymeric weight fragments.
Ru. Thus, for a given agarose gel,
The effective range of separation decreases with increasing voltage. [0005] Improved macromolecule separations are
This may be achieved by optimizing the data. example
For example, pulsed field gel electrophoresis (pulsed-field gel electrophoresis)
field gel electrophoresis
(PFGE)) is a recent advance, but given
allow greater degradation of DNA fragments with respect to the applied voltage.
Ru. Schwartz and Canter
ntor) is applied to two electric fields pulsating alternately at about 90°.
Large DNA in selectively subjected agarose gel
It teaches that it is possible to disassemble fragments. (
Schwartz, D. , and Cantor,
C. , ”Separation of Yeast
Chromosome-sized DNAs by
Pulsed-field Gradient Ge
l Electrophoresis” Cell 3
7:67, 1984) forcing molecules to change direction between pulses.
By redirecting the DNA, longer DNA fragments are
can be separated from shorter fragments. However, the induced electric field
There is no uniformity throughout the gel using this technique. huge
molecules depend on where the sample is placed in the gel
tend to move through the gel with a specific mobility and trajectory.
Ru. This position-dependent distortion
dent distortion) is the electric field imbalance.
Due to uniformity, the lane-to-lane of the band
lane). [0006] Carle et al.
A simple modification, field inversion gel electrophoresis (F
ield-inversion gel electr
ophoresis (FIGE)) is described.
, in which the alternating fields are 180° apart. So
The polarity of has a forward pulse time longer than the reverse pulse time
the net forward sun.
Pull migration (net forward sa
sample migration). this technology
is 800 kb (kilobases)
provides an acceptable decomposition. Sections exceeding a certain size
For fragments, however, large DNA molecules and
Small DNA molecules tend to move together. electrophoresis
Gradually changing (grading) the voltage across the course of the
increasing the separation distance before this occurs by
Can be done. However, if the size of the molecules to be separated is
A set of parameters must be selected first. (
Carle, G. , Frank, M. , and
Olson, M. , ”Electrophore
tic Separations of Large
DNA Molecules by Periodi
c Conversion of the Elect
ric Field,” Science 232:
65, 1986) [0007] Gardiner
(Gardiner) and Patterson (Patterso)
n), the gel is oriented vertically and the usual 4-electrode array is
transverse bars placed forward and rearward rather than in the plane of
- alternating field electrophoresis (tran
sversealternating field e
electrophoresis (TAFE) system
It describes the system. (Gardiner, K., L.
aas, W. , and Patterson, D.
．． , ”Fraction of Large
e Mammalian DNA Restrictio
n Fragments Using Vertica
l Pulsed-Field Gradient G
el Electrophoresis,” Som.
Cell Molecular Genetics 1
2: 185, 1986) Chu (C
hu) et al. have shown that the decisive factor in decomposing large DNA fragments is
The main factor is the difference between two directions of an alternating electric field greater than 90°.
It teaches that it is important to maintain the angle. (Ch.
u, G. , Vollrath, D. , and
Davis, R. , “Separation of
LargeDNA Molecules by Co
Tour-clamped Homogeneous
Electric Fields,” Science
e 234: 1582, 1986) Contour fixed homogeneity
The electric field or CHEF system was placed at 120°
A hexagonal array with pairs of electrodes is used. Ideal for electrodes
We use a transistor to force a voltage that is equal to
Fixed. The increasing decomposition causes the sample's coma gray
obtained without co-migration. CHEF-DR II (BIO-RAD, Richmond
(California) uses this technology to
It is widely used for the separation of A molecules. (U.S. patent
No. 4878008, Contour-clamped
homogeneous electric gen
erator, C. [Problem to be solved by the invention] However, the current
The system allows users to set parameters for electrophoresis operations.
must be determined and entered manually. this
The technology is inefficient and requires a lot of trial and error by the user.
request. In particular, this approach can be used under various configuration conditions.
can be assembled from electrophoretic separation of DNA standards in
knowledge base is not available. The present invention covers these and
Provide new technology and equipment to meet other requirements. [Means and effects for solving the problems] The present invention provides
Improving electrophoretic separation of macromolecules with the help of computers
We provide a system for This system separates
known under various planning conditions chosen to optimize
Separate macromolecules of specific DNA size
knowledge derived from calibration data obtained by
Stay on base. In the preferred embodiment, the knowledge base
The process uses the CHEF mapper program (Mapper P
constants, functions, and tables as part of
It will be materialized in In operation, the user selects the smallest
Enter the DNA size and maximum DNA size. This feeling
Based on the information, the system is designed to
Onch time, final switch time, forward voltage gradient, and reverse
Automatically configure electrophoresis equipment using voltage gradient parameters
calibrate to, thereby increasing electrophoretic resolution. EXAMPLES Overview The present invention provides improvements in the separation and purification of macromolecules, particularly DNA.
methods and apparatus which work in conjunction with electrophoretic means to
provide. According to the invention, users should be separated
Enter information about the macromolecule sample. Maximize separation
The parameters to be optimized are determined then. Finally, electricity
The migration means is calibrated using these parameters. With respect to FIG. 1, the present invention
TEM 150, CHEF Mapper 120, and electrophoresis
Such as the system 100 comprising the device or means 130
It can be realized in a system that computer system
150 further includes a central processor 108, a system memory
input means i.e. keyboard 103, mouse
102, fixed disk 104, display monitor 10
1, external interface 105, printer 109, and
and an input/output controller 106. computer
The components of data system 150 are connected to system bus 115.
connected, thereby providing a link between all components.
Ru. In a preferred embodiment, a suitably programmed
Used by IBM compatible personal computers
International Business
Machines, Armonk, NY),
Microsoft Windows
Windows) (Microsoft, Redmon
d, WA). With respect to FIG. 2, CHEF mapper 120 and
Also shown is a pneumophoresis device 130. mapper 1
20 is coupled to device 130 and is thus optimized
The parameters can be used to directly calibrate device 130. suitable
In some embodiments, apparatus 130 uses CHEF technology.
It is an electrophoresis means. Macromolecule samples are first placed in the wells.
(well) and move across the gel in response to an electric field.
move. More specifically, the sample is applied to multiple electrodes.
Therefore, it moves under the influence of the electric field vector generated. In one embodiment, CHEF mapper 120
You can also connect an external interface such as an RS-232 serial port.
coupled to face 105 and thereby mapper 12
0 can receive output from system 150, which then
is used to calibrate the electrophoresis apparatus 130 at. other
In the embodiment, CHEF mapper 120
121 and/or bar wand (barcode reader)
122 to its own input device, such as
The mapper 120 therefore receives an alternative input form from the user.
information directly from the format or from other systems.
can be received. As shown in FIG. 1, the mapper 120
The external interface 105 also has a keyboard 121 and
and bar wand 122, all at the same time. System 100 operates in a fully automatic mode.
do. The user selects the macromolecules expected to be separated (
Minimum and maximum fragment size (kilograms) of typically DNA)
(in kilobase). to the applicant
thus provides a knowledge base of discovered relationships, which
Therefore, the system 100 uses the optimal parameters, namely:
forward voltage gradient, reverse voltage gradient, voltage gradient, angle, initial
switch time, final switch time, and run period (R
Duration), the same as the best electrophoresis technology.
You can decide accordingly. In this way, the maximum
Separation of macromolecules can be achieved. In an alternative embodiment, system 100 is semi-autonomous.
operating in dynamic mode, the user can create agarose molds (agar
agarose type), agarose concentration (agarose type), agarose concentration (agarose
concentration), buffer type (bu
buffer type ), buffer concentration (buffer
concentration), and temperature.
Additional parameters can be specified. These values are then D
Combined with NA size, to determine the optimal parameters.
used for Individual parameters for system 100
will be described in more detail next. The forward voltage gradient is FI
Voltage difference (V) per distance used during forward pulse of GE
/cm). On the other hand, the reverse voltage gradient
is the voltage difference per distance (V/cm). Voltage
The gradient is simply the difference between the forward voltage gradient and the reverse voltage gradient. The switching time is determined by the electrophoresis device 130 (
(before switching again) how long to stay in a particular state
It is. This way, this means that a particular set of vectors (a particular
The amount of time the system maintains the voltage gradient in angle
It is a gap. The switch time is set throughout the course of electrophoresis.
It slopes (changes) depending on the direction. Preferably, the switch time is increased
Tilted in a way. More preferably, the switch time is
Slopes in a non-linear and increasing manner. In a preferred embodiment,
The switch time is logarithmically sloped. In an alternative embodiment,
The switch time slopes hyperbolically. Also for other examples
For example, if X is a normalized run period,
and Y is the difference between the initial switch time and the final switch time.
of the Cartesian coordinate system, which is the normalized difference between
slope by defining a curve on a coordinate system like
Specify. The ramping technique used
chnique ), the switch time is
The initial switch time and the upper and lower limits of the switch time slope.
It is preferable to divide the final switch time. [0020] Angle and run period have their usual connotations
keep. An angle is maintained between the alternating electric field and the sample.
This is the angle at which the Historically, forward or downward equals 0°
defined as the new angle, and with respect to the opposite or upward
Degrees are equal to 180°. Finally, run the electrophoresis
is performed by the electrophoresis apparatus 130 in the time interval
be. Users may be familiar with a particular set of terms.
Because of the nature of the program, three different types of programs are suitable for implementation.
Used in the examples. That is, 1) FIGE, 2
) two-state CHEF, and 3) multi-state CHEF. This grouping is useful for viewing run parameters, for example.
assist the user. FIGE is forward pulse (angle = 0°)
and has only reverse pulses (angle = 180°). multistate
In CHEF or program type, the angle is 0°.
Equal time sequential pulses occur, but also positive or negative
Some offset exists. For example, on one side
60° has a corresponding offset of -60°.
In this case, the net load is in the forward direction. 2nd condition
The condition CHEF indicates that it includes a
Same as multistate CHEF except that it uses
It's like that. Thus, for angles of 60° and -60°
For this, there will be an included angle of 120°. [0022] The three program types are subset/spread
- has a perset relationship. All program types are
The state can be described by the CHEF program type. This program
The program type may contain up to 8 blocks, each block being
It can contain up to 15 states (vectors). 2 states
The program type is one block with two symmetrical states.
including. Everything about each parameter in both vectors
The values of are the same except for the angle. Since the angle group is symmetrical,
, they sum to zero (forward). FIGE is also a special case of multistate CHEF.
This is the case. It also creates one block with two vectors.
Consisting of locks. However, these vectors are
Not a title. One vector has an angle of 0° (order),
while the other has an angle of 180° (opposite). Regarding standard conditions in fully automatic mode
, the following values are for small DNA sizes (<2500kb):
It is assumed that 1) Agarose type = “low EEO” (industrial standard), 2) A
Gallose concentration = 1.0%, 3) Buffer type = TBE (tris-borate E
DTA), 4) Buffer concentration = 0.5X TBE, or
5) Buffer temperature = 14 degrees Celsius The following standard conditions are assumed for larger DNA sizes:
Ru. 1) Agarose type = low EEO, 2) Agarose concentration = 0.8%, 3) Buffer type = TAE (tris-acetate)
EDTA ), 4) Buffer concentration = 1.0X TAE
, and 5) buffer temperature = 14 degrees Celsius. The system 100 knowledge base optimizes the separation.
Known service under various planning conditions chosen to optimize
Calibration obtained by isolating DNA molecules of
drawn from the data. In a preferred embodiment, the knowledge
Knowledgebase is available as part of the CHEF mapper program.
, embodied in constants, functions, and tables. The function is
Obtained using term regression and curve regression. The generated surface is
, a distance-weighted moving minimum smoothing performed on the normalized data
distance weighted mov
ing least squares procedure
re) is compatible. (See Lancaster, P.
eter, and Salkauskas, Cur
ve and Surface Fitting:
An Introduction, Academic
Press, 1974; McLain, Dr.
awing contours from Arbit
rary Data Points, Compute.
J. , 17: 318-324, 1974) function
and the matching surface are both interpolation
n) and extrapolation
) is used in some cases. The fully automatic operation of the DNA size effect system 100 can be performed in four different cases.
split and the actual case is the most input by the user.
Determined by large DNA size and minimum DNA size
. In a preferred embodiment, three ranges of DNA size are (
1) 1-50kb, (2) 50-2200kb, and
(3) Greater than 2500 kb. Each range
Macromolecules are available to optimize the separation
has its own specific set of properties. For DNA fragments of 1-50 kb, e.g.
For example, movement in a pulsed field depends on its size
is a logarithmic function of This is a movement that is linear with size.
This contrasts with larger DNA fragments that have multiple degrees of strength. different
Fragments of different sizes are not equally sensitive to fields of different strengths.
stomach. Larger fragments, e.g. 30-50kb,
It moves at 9V/cm relatively faster than fragments of approximately 1-25 kb.
move. Different field strengths correspond to smaller bands (10kb
large spacing (distance between DNA bands) that occurs with
can be used to reduce As a result, a more uniform
Intervals occur over the entire range. In this way,
A preferred embodiment is (compared to a pulsed field of the same voltage).
) 50 kb or less in order to spread the fragments in a more uniform manner.
using different voltages for the forward and reverse fields for the fragments of
Using field inversion techniques. [0028] DNA between 50 and 2500kb is
represents the range first described by Ruth and Cantor
. Many pulsed field gel separations have a D within this range.
Designed to separate NA. In order to separate large fragments of DNA,
Strength prevents large molecules from becoming entangled and breaking.
must be reduced in order to However, these
The low field strength of requires extremely long run times. two
The angle between the fields affects the mobility of the DNA, so the run
Time can be decreased by decreasing the angle between the fields
There is sex. System Operation The operation of system 100 will now be described in detail. Figure 3 shows
This is an overview of the CHEF mapper program 200, and the central
System memory 10 for instructing processor 108
A set of procedures or steps stored in 7. In other embodiments, the CHEF mapper program may e.g.
Stored in firmware such as ROM, the system update
mapper 120 or directly to the mapper 120.
. To start program 200, step
In step 201, the user uses keyboard 103 and
/or enter input parameters into mouse 102; If program 200 is directly coupled to mapper 120,
If so, the parameters can be entered using the keyboard 121 or
It is more preferably entered into wand 122 . into automatic mode
Regarding, the user can specify the minimum DN of the samples to be separated.
All you need to do is enter the A size and maximum DNA size.
be. For non-automatic mode, users can also
agarose concentration, buffer type, buffer concentration,
and temperature. Regardless of mode, input parameters
The data may be stored in system memory 107 and/or fixed data.
Table Input Parms placed on disk 104
is memorized. [0032] In step 202, if the input
The maximum DNA size (“High”) obtained is less than 50kb.
If less than or equal, the procedure executes case 1.
Then proceed to step 203. Otherwise, the procedure is
Proceed to step 204. If High is step 20
If less than or equal to 2500kb in 4, the hand
Continue to step 205 to execute case 2
. Otherwise, the procedure continues at step 206. if
Also High is 6000kb in step 206
is also less than or equal to, the procedure returns the input minimum D
Is the NA size (Low) smaller than 2000kb?
Proceed to step 208 to determine whether
nothing. If this is true, the routine proceeds to step 21
Proceed to execute case 4 at step 0. That's right
If not, the procedure returns case 3 in step 209.
Proceed to execute. However, if High is a step
206 seems to be less than or equal to 6000kb
If not, all output values are set to zero in step 207.
set equal. After this step or in case
After 1-4 has been executed, the procedure proceeds to step 211.
and keep all output values in the Output Parms table.
do. In step 212, these values are
directly via RAM 200 or via an external interface.
It is sent to the CHEF mapper 120 via 105.
. CHEF mapper 120 then determines the optimal angle, initial
Switch time, last switch time, voltage, run period, order
The electrophoresis device 130 is calibrated using a voltage and a reverse voltage.
Correct. In the CHEF mapper program that forms the individual case knowledge base:
The individual cases are described next. Figure 4 shows case 1, i.e.
Step when High is less than or equal to 50kb
show. In step 301, field inver-
gel electrophoresis (field-inversion)
gel electrophoresis (FIGE
)) Technology is selected. In step 302,
The voltage gradient is set equal to 9V/cm. step
303, the reverse voltage gradient is set equal to 6V/cm.
determined. In step 304, the angle is 180°.
and the run distance is set equal to 10cm.
Ru. In step 305, a switch time is determined.
Ru. FIGS. 5 and 6 show the steps for determining the switch time.
3 is a flowchart showing steps. At step 401
If Low is smaller than 25kb, then
, the initial switch time is 1 in step 402.
．． Set equal to 2 seconds. Otherwise, the procedure is
, the following relationship (function 1L) Y=0.0+0.015797X+0.001365X
Step 4 to calculate the initial switch time using
Proceed to 03. where X is the value of Low and Y is in seconds
The switch time depends on the In step 404, also
If the initial switch time is between 0.0 seconds and 0.05 seconds.
If so, the initial switch time is set to 0 in step 405.
．． Set equal to 0.05 seconds. Otherwise step
405 is skipped. In step 406,
The final switch time is determined by the following relationship (function 1U): Y=0.0+0.009166X+0.000137X
2, where X is the value of High and Y is the second
This is the switch time. The result of Y is empirically derived.
It is multiplied by the extracted adjustment coefficient of 1.15. Step 4
07, if the final switch time is 0.0 seconds and 0
．． 05 seconds, then step 408 is executed.
and the final switch time is set equal to 0.05 seconds.
. Otherwise step 408 is skipped. In step 409, if the initial switch time
If it is greater than the final switch time, then step
410 for initial and final switch times.
Values are reversed (exchanged with each other). Otherwise
, step 410 is skipped. The subroutine is
In this way, step 305 is completed and the process returns. In step 306, if Low is 1
If it is between or equal to kb and 6kb
, then in step 308 the mobility (cm/h
r) is the following relationship (function 2) Y=0.65426181-0.0838514428
X+0.0060879216X2, where X is the value for Low.
Ru. However, if Low is in the range of step 306
If not, then move in step 307.
The degree (cm/hr) is shown in Table 1 (Appendix) and the mobility averaging table.
determined by the process. FIG. 7 shows the steps of the mobility averaging routine.
shows. In step 501, the switch time range is
, determined from the initial and final switch times. Ste
In step 502, the number of repetitions N is the switch time range.
Determined by dividing the range by 5 and rounding to the nearest integer.
. In step 503, the delta switch time (Delt
a Switch Time) is the switch time range
is determined by dividing by N. In step 504
The following constants are set. That is, sum=0.0
, i=0, current switch time = initial switch time, and
Ru. In step 505, i is
tested. If i is less than or equal to N+1
If not, then the routine continues at step 506.
Then divide sum by N and stop, then
Return to step 307. Otherwise the routine will
Following step 507, the DNA size value is Low and Step 507.
Current switch time as switch time, and bilinear interpolation
Calculate the mobility from Table 1 or Table 2 (Appendix) using
Calculate. In step 508, the mobility is added to sum.
available. In step 509, the current switch time
is incremented. In step 510, i
is incremented and the procedure returns to step 505
Return to loop. Mobility averaging rule in step 307
After the chin is completed, the run time is calculated by dividing the run distance by the mobility.
determined in step 309 by multiplying by a positive coefficient.
Ru. of several factors such as water quality and laboratory techniques.
Even the slightest variation in any one of them will affect the operation of the system 100.
It has a big impact. In order to minimize these fluctuations
, users can calibrate based on their own experience with the system.
Adjust coefficients. The output of program 200 is adjusted accordingly
be done. The case 1 routine returns to step 203.
It ends with. FIG. 8 shows the steps for case 2. Ste
In step 601, the angle is set equal to 120°.
, and the run distance is set equal to 10 cm. Ste
At step 602, the voltage gradient is equal to 6V/cm.
Set. In step 603, regarding case 2,
The initial switch time is determined. Figure 9 shows the included
Show step. In step 701, Low is 24.
kb, the initial switch is performed in step 702.
The latch time has the following relationship (Function 22): Y=0.0+0.072277X-0.008314X
2 +0.000581X3, where X is Low and Y is
It's itch time. The procedure then jumps to step 706.
sample. However, if the low
1 is not smaller than 24kb, then
In step 703, Low indicates that it is smaller than 50kb.
Tested to see if it's true. If this were true
For example, in that case, in step 704, at the initial switch
is the following relationship (Function 21U) Y=-0.0701979+0.074324119X
Determined using +0.000007336094X2
be done. where X is Low and Y is the switch time
be. The routine continues at step 706. however
, if Low is larger than 50kb in step 703
If not, then in step 705
The period switch time has the following relationship (Function 21L) Y=0.95604+0.11662974X-0.0
0000574226X2. Here, X is Low and Y is
It's itch time. In step 706, if
If the period switch time is between 0.0 seconds and 0.05 seconds,
initial and final switch times in step 707;
The values for are reversed. The routine is called like this:
The output routine completes step 603 and ends. At step 604, at the final switch
The period is determined. Figure 10 shows the steps of this method.
. In step 801, if High is 80kb
If the final switch is smaller, in step 802 the final switch
Time has the following relationship (Function 21U) Y=-0.0701979+0.074324119X
Determined using +0.000007336094X2
be done. where X is High and Y is the switch time
be. However, if High
is smaller than 80kb, the procedure goes to 803.
Proceed to the next relationship (Function 20) Y=0.0+0.007605X+0.000686X
2 to calculate the final switch time. Here X is H
igh, and Y is the switch time (seconds). procedure
continues to step 804 where 1 is set at the last switch time.
．． It is multiplied by 15. In step 805, if
If the final switch time is between 0.0 seconds and 0.05 seconds
, then in step 806 the initial and final
The values for switch time are reversed. The procedure is called
The output routine completes step 604 and ends. [0041] At step 605, at the initial switch
If the time is greater than the last switch time, then
at the initial and final switch times in step 606.
The associated values are reversed. Otherwise step 606
is skipped. In step 607, case 2
The mobility with respect to is determined. FIGS. 11 and 12 are included in step 607.
It shows the process of In step 901, if
If Low is smaller than 50kb, go to step 902.
The mobility (cm/hr) has the following relationship (function 23): Y=1.0124518X-0.4094122386
determined using Here, X is Low and Y is
It is mobility. In step 903, if
is smaller than 169kb and the last switch time is 110kb
．． If it is smaller than 0, in step 904 the mobility (
cm/hr) in Table 2 (Appendix), initial and final switch.
latch time, low as DNA size, and mobility
determined using an averaging routine (Figure 7). Or is it?
Then, the procedure ends by returning. However, even if step 903 is false,
For example, the procedure checks whether Low is smaller than 2500kb.
Proceed to step 905 to test whether. Moshiko
If true, in step 906 the initial switch
Check if the time is between 110.0 and 300.0 seconds.
will be tested. If this is true, step
The mobility (cm/hr) of 907 is shown in Table 3 (Appendix), D
Using a value of Low as NA size and linear interpolation
Determined by The procedure then calls the routine
Step 607 is completed to terminate the process. However, if the known DNA size
is smaller than 2500kb in step 905.
If so, then skip steps 906 and 907.
be done. Furthermore, if step 906 is false;
Step 907 is skipped. At step 908
and Low is whether it is smaller than 1169kb.
tested to see. If this is true,
Then in step 909 the initial switch time is 1
less than 10.0 seconds and the final switch time is 110.
If greater than 0 seconds, then the procedure goes to step 9
Proceed to 10, Table 2 (Appendix), Initial and Final Switches
Low as time, DNA size, and mobility average
Calculate the mobility using the equation procedure. Then Lou
Chin returns and ends the game. However, if it is Low, step 9
If it is not smaller than 1169kb in 08, the step
Steps 909 and 910 are skipped. Also, if you
If step 909 is false, step 910 is skipped.
be done. Under these conditions, the routine proceeds to step 911
all output values before reaching 607 and ending step 607.
Set to zero. In step 608, the run time is
is equal to the calibration factor multiplied by the travel distance and divided by the mobility.
is set to Case 2 proceedings are concluded and
Step 205 is ended. FIG. 13 shows the call made in step 209 (FIG. 3).
The steps involved in the Case 3 procedure being invoked are shown. vinegar
At step 1001, the angle is equal to 106°
It is set as follows. In step 1002, the voltage
The voltage is set equal to 2V/cm. Step
In step 1003, the initial switch time is determined by the following equation:
(Function 50) Y=8.6+0.0053X is determined using. Here, X is H multiplied by 60
igh, and Y is the switch time. Step 10
04, if the initial switch time is 0.0 and 0.04.
05 seconds, the initial step is executed in step 1005.
The switch time is set equal to 0.05 seconds. That's right
If not, step 1005 is skipped. Step 1
In 006, the final switch time is the initial switch time
is set equal to In step 1007, move
The degree (cm/hr) is determined by the following relationship (Function 51): Y=0.047+0.00000251X+-1.
5610-9X2. Here X is
Low, and Y is the mobility. At step 1008
and the run distance is set equal to 3 cm. step
At 1009, the run time is determined by adding the run distance to the calibration factor.
is set equal to multiplied by the mobility. Case
3 terminates and ends step 209. FIGS. 14 and 15 show step 210 (FIG. 3
) shows the steps of case 4 called. Step 1
At 101, the angle is set equal to 106°. In step 1102, the voltage gradient is 2V/cm.
set equal. At step 1103, when running
time is set equal to the calibration factor multiplied by 72 hours.
. In step 1104, the initial switch time is 20
Set equal to minutes. In step 1105, the most
The final switch time is determined using the following relationship (function 50): Y=8.6+0.0053X. Here, X is H multiplied by 60
igh, and Y is the mobility. At step 1106
, the final switch time is multiplied by 1.15. vinegar
In step 1107, if the final switch time is 0
．． If between 0 and 0.05 seconds, then step 1
At 108, the final switch time is equal to 0.05 seconds.
is set. Otherwise, step 1108
Kipped. In step 1109, block 1
The output value related to is stored. In steps 1110-1111, the angle
degree is set equal to 120° and the voltage gradient is 6V/c
is set equal to m. In step 1112, the most
The final switch time is set equal to 3 minutes. step
At 1113, an initial switch time is determined. FIG. 16 shows the initial switch for case 4.
Demonstrates a method for determining time. At step 1201
If Low is smaller than 24kb, then
Sometimes in step 1202 the initial switch time is
The following relationship (Function 22) Y=0.0+0.072277X-0.008314X
2 +0.000581X3. Here, X is a low value,
Y is the switch time. The routine then steps
Continuing from 1206. However, if Low
If it is not smaller than 24kb in the file 1201, then
When , the procedure proceeds to step 1203 and Low is 5.
Test if it is less than 0kb. If this is
If true, then in step 1204 the initial
The switch time is determined by the following relationship (function 21U): Y=-0.0701979+0.074324119X
Determined using +0.000007336094X2
be done. Here, X is the low value and Y is the switch time
It is between. The procedure continues at step 1206. However,
However, if Low is 50k in step 1203
If not smaller than b, then the initial switch time
In step 1205, the following relationship (function 21L) Y=0.95604+0.11662974X-0.0
0000574226X2. Here, X is a low value,
Y is the switch time. The procedure continues at step 1206 with an initial
Determine if the switch time is between 0.0 and 0.05 seconds.
Ru. If this is true, then step 12
At 07 the initial switch time is equal to 0.05 seconds. Otherwise step 1207 is skipped. vinegar
In step 1208, the initial switch time is the final switch time.
Determine whether the interval is greater than or equal to. If this is true
If so, in step 1209 the initial and final steps are
switch time is reversed. The routine ends and
Step 1113 is ended. In step 1114, the mobility (cm
/hr) is the same as mentioned for case 2.
(Fig. 11, Fig. 12). Step 1115
In , the run distance is set equal to 4 cm. Ste
In step 1116, the run time is calculated by adding the run distance to the calibration factor.
It is determined by multiplying the separation and dividing by the mobility. procedure
terminates by returning. [0053] The present invention is described in one preferred embodiment and in certain alternatives.
Although the invention has been described in detail in relation to
and are not limited to substitutes. The true scope of the invention is
, limited not by the foregoing description but by the claims.
It will be done. Effects of the Invention As explained above, according to the present invention,
Computer-assisted electrophoretic separation of macromolecules
A system for improving is provided. [Appendix] Table 1 −−−−−−−−−−−−−−−−−−−
-------------The first column (X) is the minimum DN
It is A size [Low] (kilogram base). The second column (Y) is the switch time (seconds). The third column (Z) is the electrophoretic mobility (cm/hr)
be. Data The first column (X) is the minimum DNA size [Low] (key
Robase). The second column (Y) is the switch time (seconds). The third column (Z) is electrophoretic mobility. Fitted Surface 0057
】X
YZ8.140
0e+000 2.0000e-001
2.7129e-001 8.1400e
+000 2.3000e-001
2.8031e-001 8.1400e+0
00 2.6000e-001
2.8722e-001 8.1400e+000
2.9000e-001
2.9268e-001 8.1400e+000
3.2000e-001 2.
9759e-001 8.1400e+000
3.5000e-001 3.02
69e-001 8.1400e+000
3.8000e-001 3.0842
e-001 8.1400e+000 4
．． 1000e-001 3.1486e-
001 8.1400e+000 4.4
000e-001 3.2185e-00
1 8.1400e+000 4.700
0e-001 3.2911e-001
8.1400e+000 5.0000e
-001 3.3611e-001 8.
1400e+000 5.3000e-0
01 3.4220e-001 8.14
00e+000 5.6000e-001
3.4727e-001 8.1400
e+000 5.9000e-001
3.5142e-001 8.1400e+
000 6.2000e-001
3.5486e-001 8.1400e+00
0 6.5000e-001
3.5794e-001 8.1400e+000
6.8000e-001 3
．． 6108e-001 8.1400e+000
7.1000e-001 3.6
467e-001 8.1400e+000
7.4000e-001 3.690
3e-001 8.1400e+000
7.7000e-001 3.7423e
-001 8.1400e+000 8.
0000e-001 3.7958e-0
01 0058 1.0158e+001 2.0000e
-001 2.4379e-001 1.
0158e+001 2.3000e-0
01 2.5399e-001 1.01
58e+001 2.6000e-001
2.6236e-001 1.0158
e+001 2.9000e-001
2.6923e-001 1.0158e+
001 3.2000e-001
2.7539e-001 1.0158e+00
1 3.5000e-001
2.8155e-001 1.0158e+001
3.8000e-001 2
．． 8814e-001 1.0158e+001
4.1000e-001 2.9
523e-001 1.0158e+001
4.4000e-001 3.026
4e-001 1.0158e+001
4.7000e-001 3.1004e
-001 1.0158e+001 5.
0000e-001 3.1720e-0
01 1.0158e+001 5.30
00e-001 3.2401e-001
1.0158e+001 5.6000
e-001 3.3004e-001 1
．． 0158e+001 5.9000e-
001 3.3519e-001 1.0
158e+001 6.2000e-00
1 3.3968e-001 1.015
8e+001 6.5000e-001
3.4383e-001 1.0158e
+001 6.8000e-001
3.4795e-001 1.0158e+0
01 7.1000e-001
3.5227e-001 1.0158e+001
7.4000e-001
3.5690e-001 1.0158e+001
7.7000e-001 3.
6177e-001 1.0158e+001
8.0000e-001 3.66
91e-001 0059] 1.2176e+001 2.0000e
-001 2.1715e-001 1.
2176e+001 2.3000e-0
01 2.2861e-001 1.21
76e+001 2.6000e-001
2.3852e-001 1.2176
e+001 2.9000e-001
2.4691e-001 1.2176e+
001 3.2000e-001
2.5435e-001 1.2176e+00
1 3.5000e-001
2.6154e-001 1.2176e+001
3.8000e-001 2
．． 6893e-001 1.2176e+001
4.1000e-001 2.7
663e-001 1.2176e+001
4.4000e-001 2.845
3e-001 1.2176e+001
4.7000e-001 2.9237e
-001 1.2176e+001 5.
0000e-001 2.9990e-0
01 1.2176e+001 5.30
00e-001 3.0694e-001
1.2176e+001 5.6000
e-001 3.1339e-001 1
．． 2176e+001 5.9000e-
001 3.1930e-001 1.2
176e+001 6.2000e-00
1 3.2481e-001 1.217
6e+001 6.5000e-001
3.3013e-001 1.2176e
+001 6.8000e-001
3.3540e-001 1.2176e+0
01 7.1000e-001
3.4071e-001 1.2176e+001
7.4000e-001
3.4605e-001 1.2176e+001
7.7000e-001 3.
5132e-001 1.2176e+001
8.0000e-001 3.56
88e-001 0060] 1.4194e+001 2.0000e
-001 1.9194e-001 1.
4194e+001 2.3000e-0
01 2.0479e-001 1.41
94e+001 2.6000e-001
2.1642e-001 1.4194
e+001 2.9000e-001
2.2635e-001 1.4194e+
001 3.2000e-001
2.3494e-001 1.4194e+00
1 3.5000e-001
2.4292e-001 1.4194e+001
3.8000e-001 2
．． 5084e-001 1.4194e+001
4.1000e-001 2.5
893e-001 1.4194e+001
4.4000e-001 2.672
5e-001 1.4194e+001
4.7000e-001 2.7564e
-001 1.4194e+001 5.
0000e-001 2.8362e-0
01 1.4194e+001 5.30
00e-001 2.9064e-001
1.4194e+001 5.6000
e-001 2.9722e-001 1
．． 4194e+001 5.9000e-
001 3.0374e-001 1.4
194e+001 6.2000e-00
1 3.1026e-001 1.419
4e+001 6.5000e-001
3.1678e-001 1.4194e
+001 6.8000e-001
3.2333e-001 1.4194e+0
01 7.1000e-001
3.2989e-001 1.4194e+001
7.4000e-001
3.3643e-001 1.4194e+001
7.7000e-001 3.
4295e-001 1.4194e+001
8.0000e-001 3.49
60e-001 0061] 1.6212e+001 2.0000e
-001 1.6937e-001 1.
6212e+001 2.3000e-0
01 1.8355e-001 1.62
12e+001 2.6000e-001
1.9689e-001 1.6212
e+001 2.9000e-001
2.0824e-001 1.6212e+
001 3.2000e-001
2.1764e-001 1.6212e+00
1 3.5000e-001
2.2594e-001 1.6212e+001
3.8000e-001 2
．． 3394e-001 1.6212e+001
4.1000e-001 2.4
202e-001 1.6212e+001
4.4000e-001 2.502
9e-001 1.6212e+001
4.7000e-001 2.5870e
-001 1.6212e+001 5.
0000e-001 2.6677e-0
01 1.6212e+001 5.30
00e-001 2.7406e-001
1.6212e+001 5.6000
e-001 2.8112e-001 1
．． 6212e+001 5.9000e-
001 2.8842e-001 1.6
212e+001 6.2000e-00
1 2.9598e-001 1.621
2e+001 6.5000e-001
3.0371e-001 1.6212e
+001 6.8000e-001
3.1154e-001 1.6212e+0
01 7.1000e-001
3.1938e-001 1.6212e+001
7.4000e-001
3.2717e-001 1.6212e+001
7.7000e-001 3.
3486e-001 1.6212e+001
8.0000e-001 3.42
40e-001 0062] 1.8230e+001 2.0000e
-001 1.5044e-001 1.
8230e+001 2.3000e-0
01 1.6570e-001 1.82
30e+001 2.6000e-001
1.8059e-001 1.8230
e+001 2.9000e-001
1.9314e-001 1.8230e+
001 3.2000e-001
2.0283e-001 1.8230e+00
1 3.5000e-001
2.1081e-001 1.8230e+001
3.8000e-001 2
．． 1834e-001 1.8230e+001
4.1000e-001 2.2
594e-001 1.8230e+001
4.4000e-001 2.336
8e-001 1.8230e+001
4.7000e-001 2.4146e
-001 1.8230e+001 5.
0000e-001 2.4922e-0
01 1.8230e+001 5.30
00e-001 2.5704e-001
1.8230e+001 5.6000
e-001 2.6504e-001 1
．． 8230e+001 5.9000e-
001 2.7333e-001 1.8
230e+001 6.2000e-00
1 2.8195e-001 1.823
0e+001 6.5000e-001
2.9082e-001 1.8230e
+001 6.8000e-001
2.9982e-001 1.8230e+0
01 7.1000e-001
3.0883e-001 1.8230e+001
7.4000e-001
3.1773e-001 1.8230e+001
7.7000e-001 3.
2641e-001 1.8230e+001
8.0000e-001 3.34
72e-001 0063] 2.0248e+001 2.0000e
-001 1.3526e-001 2.
0248e+001 2.3000e-0
01 1.5140e-001 2.02
48e+001 2.6000e-001
1.6750e-001 2.0248
e+001 2.9000e-001
1.8085e-001 2.0248e+
001 3.2000e-001
1.9028e-001 2.0248e+00
1 3.5000e-001
1.9738e-001 2.0248e+001
3.8000e-001 2
．． 0406e-001 2.0248e+001
4.1000e-001 2.1
092e-001 2.0248e+001
4.4000e-001 2.179
3e-001 2.0248e+001
4.7000e-001 2.2507e
-001 2.0248e+001 5.
0000e-001 2.3254e-0
01 2.0248e+001 5.30
00e-001 2.4068e-001
2.0248e+001 5.6000
e-001 2.4943e-001 2
．． 0248e+001 5.9000e-
001 2.5864e-001 2.0
248e+001 6.2000e-00
1 2.6823e-001 2.024
8e+001 6.5000e-001
2.7808e-001 2.0248e
+001 6.8000e-001
2.8806e-001 2.0248e+0
01 7.1000e-001
2.9805e-001 2.0248e+001
7.4000e-001
3.0790e-001 2.0248e+001
7.7000e-001 3.
1746e-001 2.0248e+001
8.0000e-001 3.26
46e-001 0064] 2.2266e+001 2.0000e
-001 1.2491e-001 2.
2266e+001 2.3000e-0
01 1.4088e-001 2.22
66e+001 2.6000e-001
1.5667e-001 2.2266
e+001 2.9000e-001
1.6970e-001 2.2266e+
001 3.2000e-001
1.7860e-001 2.2266e+00
1 3.5000e-001
1.8493e-001 2.2266e+001
3.8000e-001 1
．． 9091e-001 2.2266e+001
4.1000e-001 1.9
707e-001 2.2266e+001
4.4000e-001 2.033
5e-001 2.2266e+001
4.7000e-001 2.0988e
-001 2.2266e+001 5.
0000e-001 2.1711e-0
01 2.2266e+001 5.30
00e-001 2.2534e-001
2.2266e+001 5.6000
e-001 2.3461e-001 2
．． 2266e+001 5.9000e-
001 2.4457e-001 2.2
266e+001 6.2000e-00
1 2.5490e-001 2.226
6e+001 6.5000e-001
2.6547e-001 2.2266e
+001 6.8000e-001
2.7617e-001 2.2266e+0
01 7.1000e-001
2.8689e-001 2.2266e+001
7.4000e-001
2.9749e-001 2.2266e+001
7.7000e-001 3.
0776e-001 2.2266e+001
8.0000e-001 3.17
32e-001 0065] 2.4284e+001 2.0000e
-001 1.1872e-001 2.
4284e+001 2.3000e-0
01 1.3323e-001 2.42
84e+001 2.6000e-001
1.4694e-001 2.4284
e+001 2.9000e-001
1.5821e-001 2.4284e+
001 3.2000e-001
1.6653e-001 2.4284e+00
1 3.5000e-001
1.7291e-001 2.4284e+001
3.8000e-001 1
．． 7873e-001 2.4284e+001
4.1000e-001 1.8
449e-001 2.4284e+001
4.4000e-001 1.901
6e-001 2.4284e+001
4.7000e-001 1.9584e
-001 2.4284e+001 5.
0000e-001 2.0242e-0
01 2.4284e+001 5.30
00e-001 2.1100e-001
2.4284e+001 5.6000
e-001 2.2091e-001 2
．． 4284e+001 5.9000e-
001 2.3134e-001 2.4
284e+001 6.2000e-00
1 2.4208e-001 2.428
4e+001 6.5000e-001
2.5303e-001 2.4284e
+001 6.8000e-001
2.6411e-001 2.4284e+0
01 7.1000e-001
2.7523e-001 2.4284e+001
7.4000e-001
2.8625e-001 2.4284e+001
7.7000e-001 2.
9696e-001 2.4284e+001
8.0000e-001 3.06
91e-001 0066] 2.6302e+001 2.0000e
-001 1.1480e-001 2.
6302e+001 2.3000e-0
01 1.2723e-001 2.63
02e+001 2.6000e-001
1.3835e-001 2.6302
e+001 2.9000e-001
1.4760e-001 2.6302e+
001 3.2000e-001
1.5520e-001 2.6302e+00
1 3.5000e-001
1.6171e-001 2.6302e+001
3.8000e-001 1
．． 6764e-001 2.6302e+001
4.1000e-001 1.7
329e-001 2.6302e+001
4.4000e-001 1.788
1e-001 2.6302e+001
4.7000e-001 1.8440e
-001 2.6302e+001 5.
0000e-001 1.9087e-0
01 2.6302e+001 5.30
00e-001 1.9924e-001
2.6302e+001 5.6000
e-001 2.0889e-001 2
．． 6302e+001 5.9000e-
001 2.1913e-001 2.6
302e+001 6.2000e-00
1 2.2979e-001 2.630
2e+001 6.5000e-001
2.4074e-001 2.6302e
+001 6.8000e-001
2.5188e-001 2.6302e+0
01 7.1000e-001
2.6306e-001 2.6302e+001
7.4000e-001
2.7413e-001 2.6302e+001
7.7000e-001 2.
8489e-001 2.6302e+001
8.0000e-001 2.94
97e-001 0067] 2.8320e+001 2.0000e
-001 1.1212e-001 2.
8320e+001 2.3000e-0
01 1.2265e-001 2.83
20e+001 2.6000e-001
1.3138e-001 2.8320
e+001 2.9000e-001
1.3849e-001 2.8320e+
001 3.2000e-001
1.4532e-001 2.8320e+00
1 3.5000e-001
1.5190e-001 2.8320e+001
3.8000e-001 1
．． 5779e-001 2.8320e+001
4.1000e-001 1.6
332e-001 2.8320e+001
4.4000e-001 1.689
6e-001 2.8320e+001
4.7000e-001 1.7516e
-001 2.8320e+001 5.
0000e-001 1.8209e-0
01 2.8320e+001 5.30
00e-001 1.8965e-001
2.8320e+001 5.6000
e-001 1.9819e-001 2
．． 8320e+001 5.9000e-
001 2.0770e-001 2.8
320e+001 6.2000e-00
1 2.1791e-001 2.832
0e+001 6.5000e-001
2.2859e-001 2.8320e
+001 6.8000e-001
2.3952e-001 2.8320e+0
01 7.1000e-001
2.5052e-001 2.8320e+001
7.4000e-001
2.6142e-001 2.8320e+001
7.7000e-001 2.
7204e-001 2.8320e+001
8.0000e-001 2.82
11e-001 0068] 3.0338e+001 2.0000e
-001 1.1022e-001 3.
0338e+001 2.3000e-0
01 1.1966e-001 3.03
38e+001 2.6000e-001
1.2721e-001 3.0338
e+001 2.9000e-001
1.3283e-001 3.0338e+
001 3.2000e-001
1.3844e-001 3.0338e+00
1 3.5000e-001
1.4414e-001 3.0338e+001
3.8000e-001 1
．． 4922e-001 3.0338e+001
4.1000e-001 1.5
416e-001 3.0338e+001
4.4000e-001 1.595
4e-001 3.0338e+001
4.7000e-001 1.6574e
-001 3.0338e+001 5.
0000e-001 1.7260e-0
01 3.0338e+001 5.30
00e-001 1.7974e-001
3.0338e+001 5.6000
e-001 1.8762e-001 3
．． 0338e+001 5.9000e-
001 1.9651e-001 3.0
338e+001 6.2000e-00
1 2.0623e-001 3.033
8e+001 6.5000e-001
2.1649e-001 3.0338e
+001 6.8000e-001
2.2707e-001 3.0338e+0
01 7.1000e-001
2.3774e-001 3.0338e+001
7.4000e-001
2.4835e-001 3.0338e+001
7.7000e-001 2.
5880e-001 3.0338e+001
8.0000e-001 2.68
83e-001 0069] 3.2356e+001 2.0000e
-001 1.0830e-001 3.
2356e+001 2.3000e-0
01 1.1739e-001 3.23
56e+001 2.6000e-001
1.2552e-001 3.2356
e+001 2.9000e-001
1.3183e-001 3.2356e+
001 3.2000e-001
1.3551e-001 3.2356e+00
1 3.5000e-001
1.3826e-001 3.2356e+001
3.8000e-001 1
．． 4147e-001 3.2356e+001
4.1000e-001 1.4
534e-001 3.2356e+001
4.4000e-001 1.500
0e-001 3.2356e+001
4.7000e-001 1.5559e
-001 3.2356e+001 5.
0000e-001 1.6197e-0
01 3.2356e+001 5.30
00e-001 1.6897e-001
3.2356e+001 5.6000
e-001 1.7670e-001 3
．． 2356e+001 5.9000e-
001 1.8526e-001 3.2
356e+001 6.2000e-00
1 1.9458e-001 3.235
6e+001 6.5000e-001
2.0443e-001 3.2356e
+001 6.8000e-001
2.1456e-001 3.2356e+0
01 7.1000e-001
2.2478e-001 3.2356e+001
7.4000e-001
2.3490e-001 3.2356e+001
7.7000e-001 2.
4477e-001 3.2356e+001
8.0000e-001 2.54
37e-001 0070] 3.4374e+001 2.0000e
-001 1.0519e-001 3.
4374e+001 2.3000e-0
01 1.1328e-001 3.43
74e+001 2.6000e-001
1.2088e-001 3.4374
e+001 2.9000e-001
1.2710e-001 3.4374e+
001 3.2000e-001
1.3004e-001 3.4374e+00
1 3.5000e-001
1.3144e-001 3.4374e+001
3.8000e-001 1
．． 3352e-001 3.4374e+001
4.1000e-001 1.3
648e-001 3.4374e+001
4.4000e-001 1.403
1e-001 3.4374e+001
4.7000e-001 1.4517e
-001 3.4374e+001 5.
0000e-001 1.5108e-0
01 3.4374e+001 5.30
00e-001 1.5789e-001
3.4374e+001 5.6000
e-001 1.6554e-001 3
．． 4374e+001 5.9000e-
001 1.7396e-001 3.4
374e+001 6.2000e-00
1 1.8297e-001 3.437
4e+001 6.5000e-001
1.9241e-001 3.4374e
+001 6.8000e-001
2.0207e-001 3.4374e+0
01 7.1000e-001
2.1176e-001 3.4374e+001
7.4000e-001
2.2134e-001 3.4374e+001
7.7000e-001 2.
3072e-001 3.4374e+001
8.0000e-001 2.40
01e-001 0071] 3.6392e+001 2.0000e
-001 1.0050e-001 3.
6392e+001 2.3000e-0
01 1.0656e-001 3.63
92e+001 2.6000e-001
1.1172e-001 3.6392
e+001 2.9000e-001
1.1606e-001 3.6392e+
001 3.2000e-001
1.1962e-001 3.6392e+00
1 3.5000e-001
1.2243e-001 3.6392e+001
3.8000e-001 1
．． 2490e-001 3.6392e+001
4.1000e-001 1.2
749e-001 3.6392e+001
4.4000e-001 1.305
7e-001 3.6392e+001
4.7000e-001 1.3449e
-001 3.6392e+001 5.
0000e-001 1.3975e-0
01 3.6392e+001 5.30
00e-001 1.4654e-001
3.6392e+001 5.6000
e-001 1.5436e-001 3
．． 6392e+001 5.9000e-
001 1.6275e-001 3.6
392e+001 6.2000e-00
1 1.7151e-001 3.639
2e+001 6.5000e-001
1.8054e-001 3.6392e
+001 6.8000e-001
1.8970e-001 3.6392e+0
01 7.1000e-001
1.9884e-001 3.6392e+001
7.4000e-001
2.0783e-001 3.6392e+001
7.7000e-001 2.
1667e-001 3.6392e+001
8.0000e-001 2.25
61e-001 0072] 3.8410e+001 2.0000e
-001 9.5101e-002 3.
8410e+001 2.3000e-0
01 9.9338e-002 3.84
10e+001 2.6000e-001
1.0269e-001 3.8410
e+001 2.9000e-001
1.0589e-001 3.8410e+
001 3.2000e-001
1.0962e-001 3.8410e+00
1 3.5000e-001
1.1328e-001 3.8410e+001
3.8000e-001 1
．． 1622e-001 3.8410e+001
4.1000e-001 1.1
875e-001 3.8410e+001
4.4000e-001 1.213
4e-001 3.8410e+001
4.7000e-001 1.2447e
-001 3.8410e+001 5.
0000e-001 1.2905e-0
01 3.8410e+001 5.30
00e-001 1.3577e-001
3.8410e+001 5.6000
e-001 1.4370e-001 3
．． 8410e+001 5.9000e-
001 1.5198e-001 3.8
410e+001 6.2000e-00
1 1.6040e-001 3.841
0e+001 6.5000e-001
1.6894e-001 3.8410e
+001 6.8000e-001
1.7756e-001 3.8410e+0
01 7.1000e-001
1.8617e-001 3.8410e+001
7.4000e-001
1.9465e-001 3.8410e+001
7.7000e-001 2.
0297e-001 3.8410e+001
8.0000e-001 2.11
49e-001 0073] 4.0428e+001 2.0000e
-001 8.9854e-002 4.
0428e+001 2.3000e-0
01 9.3041e-002 4.04
28e+001 2.6000e-001
9.5698e-002 4.0428
e+001 2.9000e-001
9.8494e-002 4.0428e+
001 3.2000e-001
1.0183e-001 4.0428e+00
1 3.5000e-001
1.0521e-001 4.0428e+001
3.8000e-001 1
．． 0811e-001 4.0428e+001
4.1000e-001 1.1
067e-001 4.0428e+001
4.4000e-001 1.132
7e-001 4.0428e+001
4.7000e-001 1.1638e
-001 4.0428e+001 5.
0000e-001 1.2077e-0
01 4.0428e+001 5.30
00e-001 1.2695e-001
4.0428e+001 5.6000
e-001 1.3429e-001 4
．． 0428e+001 5.9000e-
001 1.4200e-001 4.0
428e+001 6.2000e-00
1 1.4982e-001 4.042
8e+001 6.5000e-001
1.5772e-001 4.0428e
+001 6.8000e-001
1.6572e-001 4.0428e+0
01 7.1000e-001
1.7381e-001 4.0428e+001
7.4000e-001
1.8193e-001 4.0428e+001
7.7000e-001 1.
9007e-001 4.0428e+001
8.0000e-001 1.98
44e-001 0074] 4.2446e+001 2.0000e
-001 8.4994e-002 4.
2446e+001 2.3000e-0
01 8.7585e-002 4.24
46e+001 2.6000e-001
8.9906e-002 4.2446
e+001 2.9000e-001
9.2371e-002 4.2446e+
001 3.2000e-001
9.5143e-002 4.2446e+00
1 3.5000e-001
9.7984e-002 4.2446e+001
3.8000e-001 1
．． 0066e-001 4.2446e+001
4.1000e-001 1.0
330e-001 4.2446e+001
4.4000e-001 1.061
9e-001 4.2446e+001
4.7000e-001 1.0968e
-001 4.2446e+001 5.
0000e-001 1.1412e-0
01 4.2446e+001 5.30
00e-001 1.1969e-001
4.2446e+001 5.6000
e-001 1.2609e-001 4
．． 2446e+001 5.9000e-
001 1.3289e-001 4.2
446e+001 6.2000e-00
1 1.3984e-001 4.244
6e+001 6.5000e-001
1.4691e-001 4.2446e
+001 6.8000e-001
1.5415e-001 4.2446e+0
01 7.1000e-001
1.6164e-001 4.2446e+001
7.4000e-001
1.6939e-001 4.2446e+001
7.7000e-001 1.
7738e-001 4.2446e+001
8.0000e-001 1.85
66e-001 0075] 4.4464e+001 2.0000e
-001 8.0402e-002 4.
4464e+001 2.3000e-0
01 8.2518e-002 4.44
64e+001 2.6000e-001
8.4492e-002 4.4464
e+001 2.9000e-001
8.6564e-002 4.4464e+
001 3.2000e-001
8.8827e-002 4.4464e+00
1 3.5000e-001
9.1218e-002 4.4464e+001
3.8000e-001 9
．． 3707e-002 4.4464e+001
4.1000e-001 9.6
422e-002 4.4464e+001
4.4000e-001 9.957
8e-002 4.4464e+001
4.7000e-001 1.0335e
-001 4.4464e+001 5.
0000e-001 1.0781e-0
01 4.4464e+001 5.30
00e-001 1.1293e-001
4.4464e+001 5.6000
e-001 1.1852e-001 4
．． 4464e+001 5.9000e-
001 1.2438e-001 4.4
464e+001 6.2000e-00
1 1.3036e-001 4.446
4e+001 6.5000e-001
1.3648e-001 4.4464e
+001 6.8000e-001
1.4286e-001 4.4464e+0
01 7.1000e-001
1.4962e-001 4.4464e+001
7.4000e-001
1.5684e-001 4.4464e+001
7.7000e-001 1.
6452e-001 4.4464e+001
8.0000e-001 1.72
59e-001 0076] 4.6482e+001 2.0000e
-001 7.5959e-002 4.
6482e+001 2.3000e-0
01 7.7601e-002 4.64
82e+001 2.6000e-001
7.9176e-002 4.6482
e+001 2.9000e-001
8.0843e-002 4.6482e+
001 3.2000e-001
8.2690e-002 4.6482e+00
1 3.5000e-001
8.4756e-002 4.6482e+001
3.8000e-001 8
．． 7101e-002 4.6482e+001
4.1000e-001 8.9
849e-002 4.6482e+001
4.4000e-001 9.312
7e-002 4.6482e+001
4.7000e-001 9.6985e
-002 4.6482e+001 5.
0000e-001 1.0136e-0
01 4.6482e+001 5.30
00e-001 1.0612e-001
4.6482e+001 5.6000
e-001 1.1109e-001 4
．． 6482e+001 5.9000e-
001 1.1613e-001 4.6
482e+001 6.2000e-00
1 1.2120e-001 4.648
2e+001 6.5000e-001
1.2639e-001 4.6482e
+001 6.8000e-001
1.3188e-001 4.6482e+0
01 7.1000e-001
1.3785e-001 4.6482e+001
7.4000e-001
1.4442e-001 4.6482e+001
7.7000e-001 1.
5161e-001 4.6482e+001
8.0000e-001 1.59
34e-001 0077] 4.8500e+001 2.0000e
-001 7.1639e-002 4.
8500e+001 2.3000e-0
01 7.2801e-002 4.85
00e+001 2.6000e-001
7.3961e-002 4.8500
e+001 2.9000e-001
7.5246e-002 4.8500e+
001 3.2000e-001
7.6755e-002 4.8500e+00
1 3.5000e-001
7.8571e-002 4.8500e+001
3.8000e-001 8
．． 0784e-002 4.8500e+001
4.1000e-001 8.3
490e-002 4.8500e+001
4.4000e-001 8.675
0e-002 4.8500e+001
4.7000e-001 9.0536e
-002 4.8500e+001 5.
0000e-001 9.4723e-0
02 4.8500e+001 5.30
00e-001 9.9123e-002
4.8500e+001 5.6000
e-001 1.0356e-001 4
．． 8500e+001 5.9000e-
001 1.0791e-001 4.8
500e+001 6.2000e-00
1 1.1220e-001 4.850
0e+001 6.5000e-001
1.1657e-001 4.8500e
+001 6.8000e-001
1.2123e-001 4.8500e+0
01 7.1000e-001
1.2643e-001 4.8500e+001
7.4000e-001
1.3232e-001 4.8500e+001
7.7000e-001 1.
3894e-001 4.8500e+001
8.0000e-001 1.46
21e-001 0078 Table 2 −−−−−−−−−−−−−−−−−−−−−−−−
-------------The first column (X) is the minimum DNA
Size [Low] (kilogram base). The second column (Y) is the switch time (seconds). The third column (Z) is electrophoretic mobility. Data [0080] The first column (X) is the minimum DNA size [Low] (key
Robase). The second column (Y) is the switch time (seconds). The third column (Z) is electrophoretic mobility. Compatible surface 0081
YZ4.850
0e+001 5.0000e+000
2.1572e-001 4.8500e
+001 1.0750e+001
2.4990e-001 4.8500e+0
01 1.6500e+001
2.7524e-001 4.8500e+001
2.2250e+001
2.8992e-001 4.8500e+001
2.8000e+001 3.
0062e-001 4.8500e+001
3.3750e+001 3.07
61e-001 4.8500e+001
3.9500e+001 3.0930
e-001 4.8500e+001 4
．． 5250e+001 3.0986e-
001 4.8500e+001 5.1
000e+001 3.1237e-00
1 4.8500e+001 5.675
0e+001 3.1459e-001
4.8500e+001 6.2500e
+001 3.1518e-001 4.
8500e+001 6.8250e+0
01 3.1519e-001 4.85
00e+001 7.4000e+001
3.1607e-001 4.8500
e+001 7.9750e+001
3.1763e-001 4.8500e+
001 8.5500e+001
3.1890e-001 4.8500e+00
1 9.1250e+001
3.1923e-001 4.8500e+001
9.7000e+001 3
．． 1973e-001 4.8500e+001
1.0275e+002 3.2
037e-001 4.8500e+001
1.0850e+002 3.191
8e-001 4.8500e+001
1.1425e+002 3.1534e
-001 4.8500e+001 1.
2000e+002 3.0926e-0
01 0082 9.1825e+001 5.0000e
+000 1.8147e-001 9.
1825e+001 1.0750e+0
01 2.2043e-001 9.18
25e+001 1.6500e+001
2.4968e-001 9.1825
e+001 2.2250e+001
2.6541e-001 9.1825e+
001 2.8000e+001
2.7712e-001 9.1825e+00
1 3.3750e+001
2.8517e-001 9.1825e+001
3.9500e+001 2
．． 8790e-001 9.1825e+001
4.5250e+001 2.9
026e-001 9.1825e+001
5.1000e+001 2.953
3e-001 9.1825e+001
5.6750e+001 2.9984e
-001 9.1825e+001 6.
2500e+001 3.0209e-0
01 9.1825e+001 6.82
50e+001 3.0310e-001
9.1825e+001 7.4000
e+001 3.0507e-001 9
．． 1825e+001 7.9750e+
001 3.0742e-001 9.1
825e+001 8.5500e+00
1 3.0899e-001 9.182
5e+001 9.1250e+001
3.0997e-001 9.1825e
+001 9.7000e+001
3.1170e-001 9.1825e+0
01 1.0275e+002
3.1344e-001 9.1825e+001
1.0850e+002
3.1302e-001 9.1825e+001
1.1425e+002 3.
0986e-001 9.1825e+001
1.2000e+002 3.04
48e-001 0083] 1.3515e+002 5.0000e
+000 1.5246e-001 1.
3515e+002 1.0750e+0
01 1.9044e-001 1.35
15e+002 1.6500e+001
2.2009e-001 1.3515
e+002 2.2250e+001
2.3880e-001 1.3515e+
002 2.8000e+001
2.5327e-001 1.3515e+00
2 3.3750e+001
2.6321e-001 1.3515e+002
3.9500e+001 2
．． 6808e-001 1.3515e+002
4.5250e+001 2.7
284e-001 1.3515e+002
5.1000e+001 2.796
8e-001 1.3515e+002
5.6750e+001 2.8586e
-001 1.3515e+002 6.
2500e+001 2.8994e-0
01 1.3515e+002 6.82
50e+001 2.9233e-001
1.3515e+002 7.4000
e+001 2.9471e-001 1
．． 3515e+002 7.9750e+
001 2.9725e-001 1.3
515e+002 8.5500e+00
1 2.9905e-001 1.351
5e+002 9.1250e+001
3.0058e-001 1.3515e
+002 9.7000e+001
3.0308e-001 1.3515e+0
02 1.0275e+002
3.0533e-001 1.3515e+002
1.0850e+002
3.0552e-001 1.3515e+002
1.1425e+002 3.
0318e-001 1.3515e+002
1.2000e+002 2.98
67e-001 0084] 1.7848e+002 5.0000e
+000 1.2463e-001 1.
7848e+002 1.0750e+0
01 1.5894e-001 1.78
48e+002 1.6500e+001
1.8783e-001 1.7848
e+002 2.2250e+001
2.1054e-001 1.7848e+
002 2.8000e+001
2.2902e-001 1.7848e+00
2 3.3750e+001
2.4163e-001 1.7848e+002
3.9500e+001 2
．． 4907e-001 1.7848e+002
4.5250e+001 2.5
618e-001 1.7848e+002
5.1000e+001 2.645
8e-001 1.7848e+002
5.6750e+001 2.7215e
-001 1.7848e+002 6.
2500e+001 2.7777e-0
01 1.7848e+002 6.82
50e+001 2.8143e-001
1.7848e+002 7.4000
e+001 2.8420e-001 1
．． 7848e+002 7.9750e+
001 2.8702e-001 1.7
848e+002 8.5500e+00
1 2.8921e-001 1.784
8e+002 9.1250e+001
2.9123e-001 1.7848e
+002 9.7000e+001
2.9419e-001 1.7848e+0
02 1.0275e+002
2.9677e-001 1.7848e+002
1.0850e+002
2.9744e-001 1.7848e+002
1.1425e+002 2.
9582e-001 1.7848e+002
1.2000e+002 2.92
15e-001 0085] 2.2180e+002 5.0000e
+000 9.3584e-002 2.
2180e+002 1.0750e+0
01 1.2529e-001 2.21
80e+002 1.6500e+001
1.5495e-001 2.2180
e+002 2.2250e+001
1.8200e-001 2.2180e+
002 2.8000e+001
2.0457e-001 2.2180e+00
2 3.3750e+001
2.2010e-001 2.2180e+002
3.9500e+001 2
．． 3015e-001 2.2180e+002
4.5250e+001 2.3
953e-001 2.2180e+002
5.1000e+001 2.495
5e-001 2.2180e+002
5.6750e+001 2.5853e
-001 2.2180e+002 6.
2500e+001 2.6543e-0
01 2.2180e+002 6.82
50e+001 2.7011e-001
2.2180e+002 7.4000
e+001 2.7355e-001 2
．． 2180e+002 7.9750e+
001 2.7685e-001 2.2
180e+002 8.5500e+00
1 2.7947e-001 2.218
0e+002 9.1250e+001
2.8193e-001 2.2180e
+002 9.7000e+001
2.8525e-001 2.2180e+0
02 1.0275e+002
2.8811e-001 2.2180e+002
1.0850e+002
2.8917e-001 2.2180e+002
1.1425e+002 2.
8816e-001 2.2180e+002
1.2000e+002 2.85
26e-001 0086] 2.6512e+002 5.0000e
+000 6.2018e-002 2.
6512e+002 1.0750e+0
01 9.3450e-002 2.65
12e+002 1.6500e+001
1.2447e-001 2.6512
e+002 2.2250e+001
1.5449e-001 2.6512e+
002 2.8000e+001
1.8017e-001 2.6512e+00
2 3.3750e+001
1.9835e-001 2.6512e+002
3.9500e+001 2
．． 1100e-001 2.6512e+002
4.5250e+001 2.2
266e-001 2.6512e+002
5.1000e+001 2.344
3e-001 2.6512e+002
5.6750e+001 2.4496e
-001 2.6512e+002 6.
2500e+001 2.5313e-0
01 2.6512e+002 6.82
50e+001 2.5872e-001
2.6512e+002 7.4000
e+001 2.6291e-001 2
．． 6512e+002 7.9750e+
001 2.6678e-001 2.6
512e+002 8.5500e+00
1 2.6980e-001 2.651
2e+002 9.1250e+001
2.7268e-001 2.6512e
+002 9.7000e+001
2.7636e-001 2.6512e+0
02 1.0275e+002
2.7948e-001 2.6512e+002
1.0850e+002
2.8090e-001 2.6512e+002
1.1425e+002 2.
8045e-001 2.6512e+002
1.2000e+002 2.78
24e-001 0087] 3.0845e+002 5.0000e
+000 3.2160e-002 3.
0845e+002 1.0750e+0
01 6.4833e-002 3.08
45e+002 1.6500e+001
9.6914e-002 3.0845
e+002 2.2250e+001
1.2816e-001 3.0845e+
002 2.8000e+001
1.5563e-001 3.0845e+00
2 3.3750e+001
1.7604e-001 3.0845e+002
3.9500e+001 1
．． 9140e-001 3.0845e+002
4.5250e+001 2.0
546e-001 3.0845e+002
5.1000e+001 2.190
9e-001 3.0845e+002
5.6750e+001 2.3130e
-001 3.0845e+002 6.
2500e+001 2.4079e-0
01 3.0845e+002 6.82
50e+001 2.4729e-001
3.0845e+002 7.4000
e+001 2.5228e-001 3
．． 0845e+002 7.9750e+
001 2.5676e-001 3.0
845e+002 8.5500e+00
1 2.6022e-001 3.084
5e+002 9.1250e+001
2.6352e-001 3.0845e
+002 9.7000e+001
2.6760e-001 3.0845e+0
02 1.0275e+002
2.7103e-001 3.0845e+002
1.0850e+002
2.7281e-001 3.0845e+002
1.1425e+002 2.
7288e-001 3.0845e+002
1.2000e+002 2.71
28e-001 0088] 3.5178e+002 5.0000e
+000 3.8103e-003 3.
5178e+002 1.0750e+0
01 3.7852e-002 3.51
78e+002 1.6500e+001
7.0912e-002 3.5178
e+002 2.2250e+001
1.0275e-001 3.5178e+
002 2.8000e+001
1.3090e-001 3.5178e+00
2 3.3750e+001
1.5313e-001 3.5178e+002
3.9500e+001 1
．． 7123e-001 3.5178e+002
4.5250e+001 1.8
771e-001 3.5178e+002
5.1000e+001 2.033
2e-001 3.5178e+002
5.6750e+001 2.1738e
-001 3.5178e+002 6.
2500e+001 2.2831e-0
01 3.5178e+002 6.82
50e+001 2.3579e-001
3.5178e+002 7.4000
e+001 2.4164e-001 3
．． 5178e+002 7.9750e+
001 2.4680e-001 3.5
178e+002 8.5500e+00
1 2.5074e-001 3.517
8e+002 9.1250e+001
2.5451e-001 3.5178e
+002 9.7000e+001
2.5907e-001 3.5178e+0
02 1.0275e+002
2.6291e-001 3.5178e+002
1.0850e+002
2.6510e-001 3.5178e+002
1.1425e+002 2.
6562e-001 3.5178e+002
1.2000e+002 2.64
53e-001 0089] 3.9510e+002 5.0000e
+000 0.0000e+000 3.
9510e+002 1.0750e+0
01 1.2028e-002 3.95
10e+002 1.6500e+001
4.6202e-002 3.9510
e+002 2.2250e+001
7.8499e-002 3.9510e+
002 2.8000e+001
1.0673e-001 3.9510e+00
2 3.3750e+001
1.3022e-001 3.9510e+002
3.9500e+001 1
．． 5045e-001 3.9510e+002
4.5250e+001 1.6
909e-001 3.9510e+002
5.1000e+001 1.868
9e-001 3.9510e+002
5.6750e+001 2.0309e
-001 3.9510e+002 6.
2500e+001 2.1562e-0
01 3.9510e+002 6.82
50e+001 2.2417e-001
3.9510e+002 7.4000
e+001 2.3094e-001 3
．． 9510e+002 7.9750e+
001 2.3684e-001 3.9
510e+002 8.5500e+00
1 2.4135e-001 3.951
0e+002 9.1250e+001
2.4567e-001 3.9510e
+002 9.7000e+001
2.5075e-001 3.9510e+0
02 1.0275e+002
2.5513e-001 3.9510e+002
1.0850e+002
2.5778e-001 3.9510e+002
1.1425e+002 2.
5868e-001 3.9510e+002
1.2000e+002 2.57
99e-001 0090] 4.3843e+002 5.0000e
+000 0.0000e+000 4.
3843e+002 1.0750e+0
01 0.0000e+000 4.38
43e+002 1.6500e+001
2.2665e-002 4.3843
e+002 2.2250e+001
5.5437e-002 4.3843e+
002 2.8000e+001
8.3931e-002 4.3843e+00
2 3.3750e+001
1.0788e-001 4.3843e+002
3.9500e+001 1
．． 2905e-001 4.3843e+002
4.5250e+001 1.4
937e-001 4.3843e+002
5.1000e+001 1.695
8e-001 4.3843e+002
5.6750e+001 1.8823e
-001 4.3843e+002 6.
2500e+001 2.0255e-0
01 4.3843e+002 6.82
50e+001 2.1237e-001
4.3843e+002 7.4000
e+001 2.2023e-001 4
．． 3843e+002 7.9750e+
001 2.2689e-001 4.3
843e+002 8.5500e+00
1 2.3197e-001 4.384
3e+002 9.1250e+001
2.3687e-001 4.3843e
+002 9.7000e+001
2.4249e-001 4.3843e+0
02 1.0275e+002
2.4741e-001 4.3843e+002
1.0850e+002
2.5055e-001 4.3843e+002
1.1425e+002 2.
5186e-001 4.3843e+002
1.2000e+002 2.51
56e-001 0091] 4.8175e+002 5.0000e
+000 0.0000e+000 4.
8175e+002 1.0750e+0
01 0.0000e+000 4.81
75e+002 1.6500e+001
0.0000e+000 4.8175
e+002 2.2250e+001
3.2418e-002 4.8175e+
002 2.8000e+001
6.1666e-002 4.8175e+00
2 3.3750e+001
8.5536e-002 4.8175e+002
3.9500e+001 1
．． 0691e-001 4.8175e+002
4.5250e+001 1.2
873e-001 4.8175e+002
5.1000e+001 1.515
6e-001 4.8175e+002
5.6750e+001 1.7284e
-001 4.8175e+002 6.
2500e+001 1.8908e-0
01 4.8175e+002 6.82
50e+001 2.0030e-001
4.8175e+002 7.4000
e+001 2.0936e-001 4
．． 8175e+002 7.9750e+
001 2.1687e-001 4.8
175e+002 8.5500e+00
1 2.2254e-001 4.817
5e+002 9.1250e+001
2.2801e-001 4.8175e
+002 9.7000e+001
2.3421e-001 4.8175e+0
02 1.0275e+002
2.3962e-001 4.8175e+002
1.0850e+002
2.4323e-001 4.8175e+002
1.1425e+002 2.
4503e-001 4.8175e+002
1.2000e+002 2.45
15e-001 0092 5.2507e+002 5.0000e
+000 0.0000e+000 5.
2507e+002 1.0750e+0
01 0.0000e+000 5.25
07e+002 1.6500e+001
0.0000e+000 5.2507
e+002 2.2250e+001
7.4310e-003 5.2507e+
002 2.8000e+001
3.6980e-002 5.2507e+00
2 3.3750e+001
6.1612e-002 5.2507e+002
3.9500e+001 8
．． 4377e-002 5.2507e+002
4.5250e+001 1.0
821e-001 5.2507e+002
5.1000e+001 1.333
7e-001 5.2507e+002
5.6750e+001 1.5700e
-001 5.2507e+002 6.
2500e+001 1.7513e-0
01 5.2507e+002 6.82
50e+001 1.8788e-001
5.2507e+002 7.4000
e+001 1.9832e-001 5
．． 2507e+002 7.9750e+
001 2.0676e-001 5.2
507e+002 8.5500e+00
1 2.1305e-001 5.250
7e+002 9.1250e+001
2.1912e-001 5.2507e
+002 9.7000e+001
2.2595e-001 5.2507e+0
02 1.0275e+002
2.3193e-001 5.2507e+002
1.0850e+002
2.3603e-001 5.2507e+002
1.1425e+002 2.
3822e-001 5.2507e+002
1.2000e+002 2.38
70e-001 0093] 5.6840e+002 5.0000e
+000 0.0000e+000 5.
6840e+002 1.0750e+0
01 0.0000e+000 5.68
40e+002 1.6500e+001
0.0000e+000 5.6840
e+002 2.2250e+001
0.0000e+000 5.6840e+
002 2.8000e+001
1.0962e-002 5.6840e+00
2 3.3750e+001
3.7679e-002 5.6840e+002
3.9500e+001 6
．． 3075e-002 5.6840e+002
4.5250e+001 8.8
843e-002 5.6840e+002
5.1000e+001 1.154
4e-001 5.6840e+002
5.6750e+001 1.4059e
-001 5.6840e+002 6.
2500e+001 1.6039e-0
01 5.6840e+002 6.82
50e+001 1.7495e-001
5.6840e+002 7.4000
e+001 1.8707e-001 5
．． 6840e+002 7.9750e+
001 1.9658e-001 5.6
840e+002 8.5500e+00
1 2.0361e-001 5.684
0e+002 9.1250e+001
2.1037e-001 5.6840e
+002 9.7000e+001
2.1775e-001 5.6840e+0
02 1.0275e+002
2.2424e-001 5.6840e+002
1.0850e+002
2.2879e-001 5.6840e+002
1.1425e+002 2.
3132e-001 5.6840e+002
1.2000e+002 2.32
12e-001 0094] 6.1172e+002 5.0000e
+000 0.0000e+000 6.
1172e+002 1.0750e+0
01 0.0000e+000 6.11
72e+002 1.6500e+001
0.0000e+000 6.1172
e+002 2.2250e+001
0.0000e+000 6.1172e+
002 2.8000e+001
0.0000e+000 6.1172e+00
2 3.3750e+001
1.5390e-002 6.1172e+002
3.9500e+001 4
．． 3372e-002 6.1172e+002
4.5250e+001 7.0
605e-002 6.1172e+002
5.1000e+001 9.770
4e-002 6.1172e+002
5.6750e+001 1.2337e
-001 6.1172e+002 6.
2500e+001 1.4452e-0
01 6.1172e+002 6.82
50e+001 1.6123e-001
6.1172e+002 7.4000
e+001 1.7534e-001 6
．． 1172e+002 7.9750e+
001 1.8611e-001 6.1
172e+002 8.5500e+00
1 1.9414e-001 6.117
2e+002 9.1250e+001
2.0173e-001 6.1172e
+002 9.7000e+001
2.0952e-001 6.1172e+0
02 1.0275e+002
2.1629e-001 6.1172e+002
1.0850e+002
2.2121e-001 6.1172e+002
1.1425e+002 2.
2412e-001 6.1172e+002
1.2000e+002 2.25
24e-001 0095] 6.5505e+002 5.0000e
+000 0.0000e+000 6.
5505e+002 1.0750e+0
01 0.0000e+000 6.55
05e+002 1.6500e+001
0.0000e+000 6.5505
e+002 2.2250e+001
0.0000e+000 6.5505e+
002 2.8000e+001
0.0000e+000 6.5505e+00
2 3.3750e+001
0.0000e+000 6.5505e+002
3.9500e+001 2
．． 3705e-002 6.5505e+002
4.5250e+001 5.2
252e-002 6.5505e+002
5.1000e+001 7.982
7e-002 6.5505e+002
5.6750e+001 1.0562e
-001 6.5505e+002 6.
2500e+001 1.2780e-0
01 6.5505e+002 6.82
50e+001 1.4659e-001
6.5505e+002 7.4000
e+001 1.6269e-001 6
．． 5505e+002 7.9750e+
001 1.7493e-001 6.5
505e+002 8.5500e+00
1 1.8432e-001 6.550
5e+002 9.1250e+001
1.9297e-001 6.5505e
+002 9.7000e+001
2.0112e-001 6.5505e+0
02 1.0275e+002
2.0805e-001 6.5505e+002
1.0850e+002
2.1328e-001 6.5505e+002
1.1425e+002 2.
1655e-001 6.5505e+002
1.2000e+002 2.17
97e-001 0096] 6.9838e+002 5.0000e
+000 0.0000e+000 6.
9838e+002 1.0750e+0
01 0.0000e+000 6.98
38e+002 1.6500e+001
0.0000e+000 6.9838
e+002 2.2250e+001
0.0000e+000 6.9838e+
002 2.8000e+001
0.0000e+000 6.9838e+00
2 3.3750e+001
0.0000e+000 6.9838e+002
3.9500e+001 3
．． 6517e-003 6.9838e+002
4.5250e+001 3.3
617e-002 6.9838e+002
5.1000e+001 6.201
5e-002 6.9838e+002
5.6750e+001 8.8006e
-002 6.9838e+002 6.
2500e+001 1.1087e-0
01 6.9838e+002 6.82
50e+001 1.3108e-001
6.9838e+002 7.4000
e+001 1.4867e-001 6
．． 9838e+002 7.9750e+
001 1.6251e-001 6.9
838e+002 8.5500e+00
1 1.7365e-001 6.983
8e+002 9.1250e+001
1.8366e-001 6.9838e
+002 9.7000e+001
1.9238e-001 6.9838e+0
02 1.0275e+002
1.9963e-001 6.9838e+002
1.0850e+002
2.0517e-001 6.9838e+002
1.1425e+002 2.
0867e-001 6.9838e+002
1.2000e+002 2.10
27e-001 0097] 7.4170e+002 5.0000e
+000 0.0000e+000 7.
4170e+002 1.0750e+0
01 0.0000e+000 7.41
70e+002 1.6500e+001
0.0000e+000 7.4170
e+002 2.2250e+001
0.0000e+000 7.4170e+
002 2.8000e+001
0.0000e+000 7.4170e+00
2 3.3750e+001
0.0000e+000 7.4170e+002
3.9500e+001 0
．． 0000e+000 7.4170e+002
4.5250e+001 1.5
670e-002 7.4170e+002
5.1000e+001 4.495
5e-002 7.4170e+002
5.6750e+001 7.1240e
-002 7.4170e+002 6.
2500e+001 9.4380e-0
02 7.4170e+002 6.82
50e+001 1.1497e-001
7.4170e+002 7.4000
e+001 1.3320e-001 7
．． 4170e+002 7.9750e+
001 1.4860e-001 7.4
170e+002 8.5500e+00
1 1.6173e-001 7.417
0e+002 9.1250e+001
1.7332e-001 7.4170e
+002 9.7000e+001
1.8308e-001 7.4170e+0
02 1.0275e+002
1.9107e-001 7.4170e+002
1.0850e+002
1.9703e-001 7.4170e+002
1.1425e+002 2.
0058e-001 7.4170e+002
1.2000e+002 2.02
21e-001 0098] 7.8502e+002 5.0000e
+000 0.0000e+000 7.
8502e+002 1.0750e+0
01 0.0000e+000 7.85
02e+002 1.6500e+001
0.0000e+000 7.8502
e+002 2.2250e+001
0.0000e+000 7.8502e+
002 2.8000e+001
0.0000e+000 7.8502e+00
2 3.3750e+001
0.0000e+000 7.8502e+002
3.9500e+001 0
．． 0000e+000 7.8502e+002
4.5250e+001 0.0
000e+000 7.8502e+002
5.1000e+001 2.904
0e-002 7.8502e+002
5.6750e+001 5.6001e
-002 7.8502e+002 6.
2500e+001 7.8938e-0
02 7.8502e+002 6.82
50e+001 9.8700e-002
7.8502e+002 7.4000
e+001 1.1671e-001 7
．． 8502e+002 7.9750e+
001 1.3337e-001 7.8
502e+002 8.5500e+00
1 1.4833e-001 7.850
2e+002 9.1250e+001
1.6158e-001 7.8502e
+002 9.7000e+001
1.7297e-001 7.8502e+0
02 1.0275e+002
1.8245e-001 7.8502e+002
1.0850e+002
1.8911e-001 7.8502e+002
1.1425e+002 1.
9261e-001 7.8502e+002
1.2000e+002 1.94
21e-001 0099] 8.2835e+002 5.0000e
+000 0.0000e+000 8.
2835e+002 1.0750e+0
01 0.0000e+000 8.28
35e+002 1.6500e+001
0.0000e+000 8.2835
e+002 2.2250e+001
0.0000e+000 8.2835e+
002 2.8000e+001
0.0000e+000 8.2835e+00
2 3.3750e+001
0.0000e+000 8.2835e+002
3.9500e+001 0
．． 0000e+000 8.2835e+002
4.5250e+001 0.0
000e+000 8.2835e+002
5.1000e+001 1.353
9e-002 8.2835e+002
5.6750e+001 4.1689e
-002 8.2835e+002 6.
2500e+001 6.4196e-0
02 8.2835e+002 6.82
50e+001 8.2402e-002
8.2835e+002 7.4000
e+001 9.9799e-002 8
．． 2835e+002 7.9750e+
001 1.1737e-001 8.2
835e+002 8.5500e+00
1 1.3360e-001 8.283
5e+002 9.1250e+001
1.4829e-001 8.2835e
+002 9.7000e+001
1.6186e-001 8.2835e+0
02 1.0275e+002
1.7352e-001 8.2835e+002
1.0850e+002
1.8144e-001 8.2835e+002
1.1425e+002 1.
8542e-001 8.2835e+002
1.2000e+002 1.87
46e-001 0100] 8.7168e+002 5.0000e
+000 0.0000e+000 8.
7168e+002 1.0750e+0
01 0.0000e+000 8.71
68e+002 1.6500e+001
0.0000e+000 8.7168
e+002 2.2250e+001
0.0000e+000 8.7168e+
002 2.8000e+001
0.0000e+000 8.7168e+00
2 3.3750e+001
0.0000e+000 8.7168e+002
3.9500e+001 0
．． 0000e+000 8.7168e+002
4.5250e+001 0.0
000e+000 8.7168e+002
5.1000e+001 0.000
0e+000 8.7168e+002
5.6750e+001 2.4724e
-002 8.7168e+002 6.
2500e+001 4.7216e-0
02 8.7168e+002 6.82
50e+001 6.5532e-002
8.7168e+002 7.4000
e+001 8.3563e-002 8
．． 7168e+002 7.9750e+
001 1.0154e-001 8.7
168e+002 8.5500e+00
1 1.1797e-001 8.716
8e+002 9.1250e+001
1.3372e-001 8.7168e
+002 9.7000e+001
1.4977e-001 8.7168e+0
02 1.0275e+002
1.6402e-001 8.7168e+002
1.0850e+002
1.7369e-001 8.7168e+002
1.1425e+002 1.
7917e-001 8.7168e+002
1.2000e+002 1.82
64e-001 0101] 9.1500e+002 5.0000e
+000 0.0000e+000 9.
1500e+002 1.0750e+0
01 0.0000e+000 9.15
00e+002 1.6500e+001
0.0000e+000 9.1500
e+002 2.2250e+001
0.0000e+000 9.1500e+
002 2.8000e+001
0.0000e+000 9.1500e+00
2 3.3750e+001
0.0000e+000 9.1500e+002
3.9500e+001 0
．． 0000e+000 9.1500e+002
4.5250e+001 0.0
000e+000 9.1500e+002
5.1000e+001 0.000
0e+000 9.1500e+002
5.6750e+001 4.5418e
-003 9.1500e+002 6.
2500e+001 2.7759e-0
02 9.1500e+002 6.82
50e+001 4.7768e-002
9.1500e+002 7.4000
e+001 6.7301e-002 9
．． 1500e+002 7.9750e+
001 8.5817e-002 9.1
500e+002 8.5500e+00
1 1.0198e-001 9.150
0e+002 9.1250e+001
1.1834e-001 9.1500e
+002 9.7000e+001
1.3704e-001 9.1500e+0
02 1.0275e+002
1.5430e-001 9.1500e+002
1.0850e+002
1.6588e-001 9.1500e+002
1.1425e+002 1.
7281e-001 9.1500e+002
1.2000e+002 1.77
88e-001 0102] 9.5832e+002 5.0000e
+000 0.0000e+000 9.
5832e+002 1.0750e+0
01 0.0000e+000 9.58
32e+002 1.6500e+001
0.0000e+000 9.5832
e+002 2.2250e+001
0.0000e+000 9.5832e+
002 2.8000e+001
0.0000e+000 9.5832e+00
2 3.3750e+001
0.0000e+000 9.5832e+002
3.9500e+001 0
．． 0000e+000 9.5832e+002
4.5250e+001 0.0
000e+000 9.5832e+002
5.1000e+001 0.000
0e+000 9.5832e+002
5.6750e+001 0.0000e
+000 9.5832e+002 6.
2500e+001 7.9093e-0
03 9.5832e+002 6.82
50e+001 2.9880e-002
9.5832e+002 7.4000
e+001 5.0797e-002 9
．． 5832e+002 7.9750e+
001 7.0155e-002 9.5
832e+002 8.5500e+00
1 8.7281e-002 9.583
2e+002 9.1250e+001
1.0473e-001 9.5832e
+002 9.7000e+001
1.2446e-001 9.5832e+0
02 1.0275e+002
1.4313e-001 9.5832e+002
1.0850e+002
1.5665e-001 9.5832e+002
1.1425e+002 1.
6548e-001 9.5832e+002
1.2000e+002 1.72
04e-001 0103] 1.0017e+003 5.0000e
+000 0.0000e+000 1.
0017e+003 1.0750e+0
01 0.0000e+000 1.00
17e+003 1.6500e+001
0.0000e+000 1.0017
e+003 2.2250e+001
0.0000e+000 1.0017e+
003 2.8000e+001
0.0000e+000 1.0017e+00
3 3.3750e+001
0.0000e+000 1.0017e+003
3.9500e+001 0
．． 0000e+000 1.0017e+003
4.5250e+001 0.0
000e+000 1.0017e+003
5.1000e+001 0.000
0e+000 1.0017e+003
5.6750e+001 0.0000e
+000 1.0017e+003 6.
2500e+001 0.0000e+0
00 1.0017e+003 6.82
50e+001 1.2092e-002
1.0017e+003 7.4000
e+001 3.4260e-002 1
．． 0017e+003 7.9750e+
001 5.4766e-002 1.0
017e+003 8.5500e+00
1 7.3684e-002 1.001
7e+003 9.1250e+001
9.2561e-002 1.0017e
+003 9.7000e+001
1.1232e-001 1.0017e+0
03 1.0275e+002
1.3101e-001 1.0017e+003
1.0850e+002
1.4604e-001 1.0017e+003
1.1425e+002 1.
5707e-001 1.0017e+003
1.2000e+002 1.65
37e-001 0104] 1.0450e+003 5.0000e
+000 0.0000e+000 1.
0450e+003 1.0750e+0
01 0.0000e+000 1.04
50e+003 1.6500e+001
0.0000e+000 1.0450
e+003 2.2250e+001
0.0000e+000 1.0450e+
003 2.8000e+001
0.0000e+000 1.0450e+00
3 3.3750e+001
0.0000e+000 1.0450e+003
3.9500e+001 0
．． 0000e+000 1.0450e+003
4.5250e+001 0.0
000e+000 1.0450e+003
5.1000e+001 0.000
0e+000 1.0450e+003
5.6750e+001 0.0000e
+000 1.0450e+003 6.
2500e+001 0.0000e+0
00 1.0450e+003 6.82
50e+001 0.0000e+000
1.0450e+003 7.4000
e+001 1.7765e-002 1
．． 0450e+003 7.9750e+
001 3.9637e-002 1.0
450e+003 8.5500e+00
1 6.0243e-002 1.045
0e+003 9.1250e+001
8.0373e-002 1.0450e
+003 9.7000e+001
1.0032e-001 1.0450e+0
03 1.0275e+002
1.1906e-001 1.0450e+003
1.0850e+002
1.3510e-001 1.0450e+003
1.1425e+002 1.
4793e-001 1.0450e+003
1.2000e+002 1.57
97e-001 0105] 1.0883e+003 5.0000e
+000 0.0000e+000 1.
0883e+003 1.0750e+0
01 0.0000e+000 1.08
83e+003 1.6500e+001
0.0000e+000 1.0883
e+003 2.2250e+001
0.0000e+000 1.0883e+
003 2.8000e+001
0.0000e+000 1.0883e+00
3 3.3750e+001
0.0000e+000 1.0883e+003
3.9500e+001 0
．． 0000e+000 1.0883e+003
4.5250e+001 0.0
000e+000 1.0883e+003
5.1000e+001 0.000
0e+000 1.0883e+003
5.6750e+001 0.0000e
+000 1.0883e+003 6.
2500e+001 0.0000e+0
00 1.0883e+003 6.82
50e+001 0.0000e+000
1.0883e+003 7.4000
e+001 1.2775e-003 1
．． 0883e+003 7.9750e+
001 2.4559e-002 1.0
883e+003 8.5500e+00
1 4.6608e-002 1.088
3e+003 9.1250e+001
6.7829e-002 1.0883e
+003 9.7000e+001
8.8271e-002 1.0883e+0
03 1.0275e+002
1.0735e-001 1.0883e+003
1.0850e+002
1.2423e-001 1.0883e+003
1.1425e+002 1.
3843e-001 1.0883e+003
1.2000e+002 1.50
02e-001 0106] 1.1316e+003 5.0000e
+000 0.0000e+000 1.
1316e+003 1.0750e+0
01 0.0000e+000 1.13
16e+003 1.6500e+001
0.0000e+000 1.1316
e+003 2.2250e+001
0.0000e+000 1.1316e+
003 2.8000e+001
0.0000e+000 1.1316e+00
3 3.3750e+001
0.0000e+000 1.1316e+003
3.9500e+001 0
．． 0000e+000 1.1316e+003
4.5250e+001 0.0
000e+000 1.1316e+003
5.1000e+001 0.000
0e+000 1.1316e+003
5.6750e+001 0.0000e
+000 1.1316e+003 6.
2500e+001 0.0000e+0
00 1.1316e+003 6.82
50e+001 0.0000e+000
1.1316e+003 7.4000
e+001 0.0000e+000 1
．． 1316e+003 7.9750e+
001 9.3618e-003 1.1
316e+003 8.5500e+00
1 3.2700e-002 1.131
6e+003 9.1250e+001
5.4944e-002 1.1316e
+003 9.7000e+001
7.6071e-002 1.1316e+0
03 1.0275e+002
9.5729e-002 1.1316e+003
1.0850e+002
1.1341e-001 1.1316e+003
1.1425e+002 1.
2876e-001 1.1316e+003
1.2000e+002 1.41
68e-001 0107] 1.1750e+003 5.0000e
+000 0.0000e+000 1.
1750e+003 1.0750e+0
01 0.0000e+000 1.17
50e+003 1.6500e+001
0.0000e+000 1.1750
e+003 2.2250e+001
0.0000e+000 1.1750e+
003 2.8000e+001
0.0000e+000 1.1750e+00
3 3.3750e+001
0.0000e+000 1.1750e+003
3.9500e+001 0
．． 0000e+000 1.1750e+003
4.5250e+001 0.0
000e+000 1.1750e+003
5.1000e+001 0.000
0e+000 1.1750e+003
5.6750e+001 0.0000e
+000 1.1750e+003 6.
2500e+001 0.0000e+0
00 1.1750e+003 6.82
50e+001 0.0000e+000
1.1750e+003 7.4000
e+001 0.0000e+000 1
．． 1750e+003 7.9750e+
001 0.0000e+000 1.1
750e+003 8.5500e+00
1 1.8498e-002 1.175
0e+003 9.1250e+001
4.1753e-002 1.1750e
+003 9.7000e+001
6.3685e-002 1.1750e+0
03 1.0275e+002
8.4069e-002 1.1750e+003
1.0850e+002
1.0258e-001 1.1750e+003
1.1425e+002 1.
1896e-001 1.1750e+003
1.2000e+002 1.33
08e-001 0108 Table 3 ---------------------
-------------The first column (X) is the DNA site.
(kilobase). The second column (Y) is the mobility (cm/hr). [0109]

[Brief explanation of the drawing]

FIG. 1 is a block diagram of the system of the present invention.

FIG. 2 is a block diagram of a mapper and an electrophoresis device.

FIG. 3 is a flowchart of the CHEF mapper program.

FIG. 4 is a flowchart of case 1.

FIG. 5 is a flowchart of switch time determination for case 1;

FIG. 6 is a flowchart of switch time determination for case 1;

FIG. 7 is a flowchart of a mobility averaging routine.

FIG. 8 is a flowchart of case 2.

FIG. 9 is a flowchart of initial switch time determination for case 2;

FIG. 10 is a flowchart of final switch time determination for case 2;

FIG. 11 is a flowchart of mobility determination for case 2;

FIG. 12 is a flowchart of mobility determination for case 2;

FIG. 13 is a flowchart of case 3.

FIG. 14 is a flowchart of case 4.

FIG. 15 is a flowchart of case 4.

FIG. 16 is a flowchart of initial switch time determination for case 4;

[Explanation of symbols]

100...System of the present invention 101...Display monitor 102...mouse 103...Keyboard 104...Fixed disk 105...External interface 106...I/O controller 107...System memory 108...Central processor 109...Printer 115...System bus 120…CHEF mapper 121...Keyboard 122...Bar wand 130...Electrophoresis means 150...computer system

Claims (10)

[Claims] 1. In an electrophoresis system, inputting a first size representing the smallest macromolecule to be resolved; a second size representing the largest macromolecule to be resolved;
storing a knowledge base representing calibration data obtained from separating macromolecules of known size under conditions that optimize the separation;
and the second size against the knowledge base, thereby generating a set of optimized parameters; and calibrating the system using the optimized parameters. , thereby optimizing the separation of macromolecules. 2. The testing step tests the first size and the second size against a knowledge base, thereby determining angle, run distance, voltage gradient, initial switch time, final switch time, and 2. The method of claim 1, comprising: selecting a run distance. 3. The calibration step calibrates the system for optimal separation by using the angle, the run distance, the voltage gradient, the initial switch time, the final switch time, and the run distance. 3. The method of claim 2, comprising: 4. The knowledge base storing step comprises storing a first plurality of relationships between macromolecule size and switching time; a second plurality of relationships between macromolecule size and mobility. Remember; and the smallest macromolecule size;
2. The method of claim 1, comprising: storing a third plurality of relationships between switch times and mobilities. 5. Electrophoresis means; a computer having one memory and one processor and selectively coupled to said electrophoresis means; coupled to said computer and displaying the smallest macromolecule to be resolved. input means for inputting a first size and for inputting a second size indicating the largest macromolecule to be resolved; a knowledge base for storing information representing calibration data obtained from separating macromolecules of sizes; a knowledge base coupled to said memory and configured to compare said first size and said second size; programming means for instructing said processor to test, thereby generating a set of optimized parameters; and calibrating said electrophoresis means using said optimized parameters;
means thereby optimizing the separation of macromolecules;
An electrophoresis system for improving the separation of macromolecules. 6. The programming means is coupled to the memory and instructs the processor to test the first size and the second size against the knowledge base, thereby determining the angle, run, etc. 6. The system of claim 5, comprising programming means for selecting distance, voltage gradient, initial switch time, final switch time, and run distance. 7. The means for calibrating the angle;
7. The system of claim 6, comprising means for calibrating the system for optimal separation by using the run distance, the voltage gradient, the initial switch time, the final switch time, and the run distance. . 8. The knowledge base is coupled to the memory and comprises: a first plurality of relationships between macromolecule size and switching time; a second plurality of relationships between macromolecule size and mobility; 6. The system of claim 5, comprising storage means for storing a third plurality of relationships between minimum macromolecule size, switching time, and mobility. 9. The system of claim 5, further comprising mapper means selectively coupled to said computer through an external interface for receiving said optimized parameters. 10. The system of claim 5, wherein said electrophoresis means is a CHEF electrophoresis device.