CN108841939A - Multiple digital pcr concentration measuring method and droplet type digital pcr system - Google Patents
Multiple digital pcr concentration measuring method and droplet type digital pcr system Download PDFInfo
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Abstract
The invention discloses a kind of multiple digital pcr concentration measuring method and droplet type digital pcr systems, concentration suitable for two or more target spots of the detection in a PCR response sample simultaneously, different target spots is indicated using different fluorophors, and the concentration measuring method includes：Each corresponding fluorophor, determines filtering bandwidth range, with the corresponding channel unit of determination, and detects the luminous intensity of each channel unit；According to the luminous spectrogram of fluorophor filtering bandwidth range corresponding with the fluorophor, the luminous contribution ratio of each fluorophor in channel unit is obtained；According to the luminous contribution ratio of each fluorophor in the luminous intensity of the channel unit and the channel unit, the concentration of each fluorophor is obtained.The present invention provides a kind of PCR density calculating methods, are corrected by algorithm, " the grey point " observed in channel are effectively reduced, and correct the fluorescence crosstalk of interchannel brought by multiple target spot.
Description
Technical field
The present invention relates to round pcr field, in particular to a kind of multiple digital pcr concentration measuring method and droplet type number
PCR system.
Background technique
Sample is usually separated into micro- reaction of a large amount of nanoliter volumes by current digital pcr technology in some way
Interior, then to this, reaction chamber carries out PCR cycle simultaneously slightly, after PCR amplification enters the period of saturation, takes terminal observation method area
The result for dividing each reaction chamber is negative or positive reaction.And in actual application, it usually needs while detecting in sample
Two or more target spots (amplification target), thus need (to be sent out after being stimulated in specific wave band with different fluorophors
Fluorescence out) indicate different target spots.In order to distinguish the fluorophor of different target spots, can generally use two or more
Fluorescence bands channel separately detects each fluorophor.
Ideal design is that each detection channels can only detect a kind of fluorophor.But actually fluorophor is luminous
Wave band is difficult to avoid not to be overlapped, as shown in Figure 1, the luminescence band of HEX fluorophor is almost by the hair of FAM fluorophor in figure
Optical band covers, so anyway optimizing the central wavelength and bandwidth of detection channels, each channel can detect difference
Fluorophor, this phenomenon is just called the fluorescence crosstalk of different interchannels.This crosstalk, the great disturbing reaction room of meeting is not
With the positive judgement of feminine gender in observation channel.In addition, different target spots will form competition in the indoor reaction of same reaction, thus shadow
The reaction efficiency of single target spot is rung, and then influences the terminal observation of the fluorophor of mark target spot, causes negative positive judgement
Fault.
Summary of the invention
In order to solve problems in the prior art, it the present invention provides a kind of multiple digital pcr concentration measuring method, overcomes
The accuracy of fluorophor measurement of concetration caused by terminal observation method and fluorescence crosstalk, the technical solution is as follows：
On the one hand, the present invention provides a kind of multiple digital pcr concentration measuring method, it is suitable for while detects in a PCR
The concentration of two or more target spots in response sample, different target spots is indicated using different fluorophors, described dense
Spending measurement method includes：
Each corresponding fluorophor, determines filtering bandwidth range, with the corresponding channel unit of determination, and detects each logical
The luminous intensity of road unit；
According to the luminous spectrogram of fluorophor filtering bandwidth range corresponding with the fluorophor, obtain in channel unit
The luminous contribution ratio of each fluorophor；
According to the luminous contribution ratio of each fluorophor in the luminous intensity of the channel unit and the channel unit,
Obtain the concentration of each fluorophor.
Further, each fluorophor according in the luminous intensity of the channel unit and the channel unit
Shine contribution ratio, and the concentration for obtaining each fluorophor includes：
The luminous intensity of each fluorophor is obtained by following formula：
INTENSITYi=a1i×c1+a2i×c2+......+aji×cj, wherein target spot quantity and for indicating target spot
Fluorophor number of species be j, i is channel number, INTENSITYiFor the strong light for detecting i-th obtained of channel
Degree, c1For the luminous intensity of first fluorophor, c2For the luminous intensity of second fluorophor, cjFor j-th of fluorophor
Luminous intensity, a1iFor the luminous contribution ratio of first fluorophor in i-th of channel, a2iIt is in i-th of channel
The luminous contribution ratio of two fluorophors, ajiFor the luminous contribution ratio of j-th of fluorophor in i-th of channel, 2≤i
≤j。
Further, further include after the luminous intensity for obtaining each fluorophor：
According to the relational model of the concentration of pre-established fluorophor and luminous intensity, shining for the fluorophor is obtained
The corresponding concentration of intensity.
Further, further include before determining filtering bandwidth range to fluorophor：Selection is different for corresponding mark
Target spot fluorophor, the wave-length coverage registrations of multiple fluorophors is greater than preset first threshold and/or the fluorescence
The corresponding wavelength difference of the emission peak of group is greater than preset second threshold.
Further, the fluorophor for indicating different target spots include but is not limited to FAM, TET, JOE, HEX, CY3,
TAMRA, ROX, Texas Red, LC RED640 and other fluorophors, the corresponding calibration of each fluorophor have luminous spectrogram.
Further, the luminous contribution ratio of each fluorophor in the channel unit is obtained by following steps：
The luminous spectrogram of different fluorophors is calculated in the integral of the corresponding filtering bandwidth range of the same channel unit, to make
For luminous contribution of the corresponding fluorophor in the channel unit；
By luminous contribution of one of fluorophor in the channel unit divided by fluorescence all in the channel unit
The luminous contribution summation of group obtains the fluorophor in the luminous contribution ratio of the channel unit.
Further, each fluorophor of the correspondence determines that filtering bandwidth range includes：
According to the wavelength characteristic of each fluorophor, the filter of corresponding selection different frequency bands width, the filtering dress
The frequency bandwidth set covers and uniquely covers the emission peak of corresponding fluorophor and/or the bandwidth of the filter
The coincidence wave-length coverage of the luminous spectrogram of multiple fluorophors is greater than preset third threshold value in spending；
Bandwidth with the frequency bandwidth of the filter of selection, as the corresponding channel unit of fluorophor.
Further, the luminous intensity of each channel unit of detection includes：
PCR response sample is filtered using selected filter one by one respectively；
Light-intensity test is carried out to the PCR response sample being filtered, obtains the luminous intensity of corresponding channel unit one by one.
Further, the target spot quantity in the PCR response sample is two, three or three or more.
On the other hand, the present invention provides a kind of droplet type digital pcr systems, real using measurement method as described above
Now to each droplet quantitative observation.
Technical solution bring provided by the invention has the beneficial effect that：
A. it is corrected by PCR algorithm, " the grey point " observed in channel is effectively reduced；
B. the fluorescence crosstalk of interchannel brought by multiple target spot is corrected；
C. it can be realized to multiple target spots in same PCR sample while carrying out measurement of concetration；
D. the bandwidth for not limiting filtering channel specially, so as to improve the port number of Multiple detection, considerably beyond tradition
Port number.
Detailed description of the invention
To describe the technical solutions in the embodiments of the present invention more clearly, make required in being described below to embodiment
Attached drawing is briefly described, it should be apparent that, drawings in the following description are only some embodiments of the invention, for
For those of ordinary skill in the art, without creative efforts, it can also be obtained according to these attached drawings other
Attached drawing.
Fig. 1 is the luminous spectrogram of FAM fluorophor Yu HEX fluorophor；
Fig. 2 is the flow chart of multiple digital pcr concentration measuring method provided in an embodiment of the present invention.
Specific embodiment
In order to enable those skilled in the art to better understand the solution of the present invention, below in conjunction in the embodiment of the present invention
Attached drawing, technical scheme in the embodiment of the invention is clearly and completely described, it is clear that described embodiment is only
The embodiment of a part of the invention, instead of all the embodiments.Based on the embodiments of the present invention, ordinary skill people
The model that the present invention protects all should belong in member's every other embodiment obtained without making creative work
It encloses.
It should be noted that description and claims of this specification and term " first " in above-mentioned attached drawing, "
Two " etc. be to be used to distinguish similar objects, without being used to describe a particular order or precedence order.It should be understood that using in this way
Data be interchangeable under appropriate circumstances, so as to the embodiment of the present invention described herein can in addition to illustrating herein or
Sequence other than those of description is implemented.In addition, term " includes " and " having " and their any deformation, it is intended that cover
Covering non-exclusive includes to be not necessarily limited to for example, containing the process, method of a series of steps or units, device, product or equipment
Step or unit those of is clearly listed, but may include be not clearly listed or for these process, methods, product
Or other step or units that equipment is intrinsic.
In the terminal observation data of multiple digital pcr, because of various disturbing factors, the fluorescence signal of many micro- reaction chambers
It can fall within negative and positive section, these points are commonly referred to as in the industry " grey point ".Certainly, disturbing factor source is complicated, no
It only limits and above-mentioned reason (type and form of such as sample template).Algorithm proposed by the present invention is to correct droplet type number
As observing interchannel crosstalk brought by multiple target spot in PCR, and it is competing because of reaction of the different target spots in same droplet
Strive brought interference.
In one embodiment of the invention, a kind of multiple digital pcr concentration measuring method is provided, suitable for detecting simultaneously
The concentration of two or more target spots in a PCR response sample, different target spots use different fluorophor marks
Show, as shown in Fig. 2, the concentration measuring method includes following below scheme：
S1, each corresponding fluorophor, determine filtering bandwidth range, with the corresponding channel unit of determination, and detect each
The luminous intensity of a channel unit.
In one embodiment of the invention, it is illustrated by taking the concentration measuring method of three target spots as an example, is three targets
Point three kinds of fluorophors of selection respectively represent the first target spot, the second target spot and third target spot, select the first principle of fluorophor
It is greater than preset first threshold (such as 100nm) for the wave-length coverage registration of fluorophor；In a further advantageous embodiment,
Selection fluorophor follows following second principle：The corresponding wavelength difference of the emission peak of the fluorophor is greater than preset the
Two threshold values (such as 30nm), alternatively, two above principle follows simultaneously.The reasons why the two principles and beneficial effect combine following
The selection of frequency bandwidth of filter illustrate together：
The determining filtering bandwidth range actually selects filter (filter plate or the filter of specific frequency bandwidth
Wave device), determine corresponding channel unit (with the frequency bandwidth of the filter of selection, as fluorescence with the specific frequency bandwidth
The bandwidth of the corresponding channel unit of group), as shown in Figure 1, FAM is shown as the region filling that curve 1 and horizontal axis surround, HEX is shown
The region filling surrounded for curve 2 and horizontal axis；The filter bandwidht range of first passage is indicated using A, B as the rectangle frame on vertex, with
E, F is that the rectangle frame on vertex indicates the filter bandwidht range of second channel.In traditional multiplexor design, in order to as far as possible
The interference of crosstalk is avoided, interchannel cannot be overlapped as far as possible, that is, vertex B has to be less than vertex E on wavelength, so very big
The quantity and design in channel are limited in degree.And in method provided by the present application, this limitation is not needed, is not limited specially
The bandwidth of filtering channel, so as to improve the port number of Multiple detection, considerably beyond traditional port number.Based on above-mentioned first
Principle, in the frequency bandwidth of filter multiple fluorophors shine spectrogram (being overlapped two-by-two) be overlapped wave-length coverage be greater than it is default
Third threshold value (such as 10nm), this with the luminescence band for evading overlapping fluorophor be as far as possible in the prior art exist it is obvious
Difference, in the prior art, the crosstalk that a variety of fluorophors generate in channel is channel noise, is detrimental to detect the channel
Interior main fluorophor, and in the present invention, the wave-length coverage registration of fluorophor can not be resistance for carrying out measurement of concetration
Hinder effect, reason is：In the application, a variety of fluorophor information in channel belong to effective information, and the first threshold is arranged
Value be for the ease of filtering when setting be overlapped wave-length coverage be greater than third threshold value filtered band width, and as setting third
Threshold value is the effective information of the correspondence fluorophor within the scope of this for ease of calculation, for example, to the curve within the scope of 1nm pickup
(especially close to the curve of axis) is integrated, integral result it is error bigger.
Based on the second principle, the frequency bandwidth covering of the filter and the transmitting for uniquely covering corresponding fluorophor
The corresponding wavelength of peak value, on the one hand, the information near the corresponding wavelength of the emission peak of fluorophor more can compared to low ebb region
The illuminated message of fluorophor is embodied, therefore, it is necessary to the frequency bandwidths of the filter of selection to cover corresponding fluorophor
The corresponding wavelength of emission peak；On the other hand, the corresponding fluorophor of the filter of each selection, i.e., in the every of selection
Although being related to multiple fluorophors in a frequency bandwidth range, corresponding fluorophor is main fluorophor, in order to anti-
Only confeuse the parimary with secondary (such as two peak points of covering), and the frequency bandwidth of the filter of selection is needed uniquely to cover corresponding fluorescence
The corresponding wavelength of the emission peak of group.To sum up two principles, when choosing filtering bandwidth range, it should from corresponding fluorescence
The a certain range of selection of left and right floating of group emission peak, is advisable with the peak value for not choosing adjacent.
In the prior art, as shown in table 1, the fluorophor for indicating different target spots include but is not limited to FAM, TET,
JOE, HEX, CY3, TAMRA, ROX, Texas Red, LC RED640 and other fluorophors (such as CY5, LC RED705 etc.
Deng), the corresponding calibration of each fluorophor has luminous spectrogram, it should be noted that the above is only part in the prior art is common
Fluorophor does not represent all fluorophors, can be free there are many kinds of different fluorophors from 520~800nm
Combination, also, the type selection of fluorophor is not inventive point of the invention, but the selection thought of fluorophor and filtering
The selection thought of device is that have certain innovation for the subsequent concentration for preferably detecting fluorophor.
Table 1
The luminous intensity of each channel unit of the detection is specially：In experimental fluorescence image checking, sample is reacted to PCR
Product are filtered using selected filter one by one respectively；Light-intensity test is carried out to the PCR response sample being filtered, by
One obtains the luminous intensity of corresponding channel unit.For example, light intensity is filtered and detected using the first filter, then is obtained
The luminous intensity INTENSITY of one channel unit1, and so on, detection obtains INTENSITY respectively2And INTENSITY3。
S2, the luminous spectrogram filtering bandwidth range corresponding with the fluorophor according to fluorophor, obtain channel unit
The luminous contribution ratio of interior each fluorophor.
As seen from Figure 1, the main contributions of the channel AB imaging are FAM fluorophor luminous spectrum, while also containing HEX fluorescence
The contribution of group luminous spectrum；The main contributions of the channel EF imaging are HEX fluorophor luminous spectrum, while containing FAM fluorescent base
The contribution of group's luminous spectrum, the i.e. size of fluorophor contribution, it is not only related with luminous spectrum, but also have with the bandwidth of filter
It closes.
In the image checking in the channel AB, the contribution of FAM fluorophor is its luminous spectrum in AB path filter bandwidth model
The integral of (AB) in enclosing, is expressed as famInAB, and the contribution of HEX fluorophor is its luminous spectrum in AB path filter bandwidth model
The integral of (AB), is expressed as hexInAB in enclosing.In the image checking in the channel EF, the contribution of FAM fluorophor is its luminous spectrum
The integral of (EF) in EF path filter bandwidth range, is expressed as famInEF, and the contribution of HEX fluorophor is its luminous spectrum
The integral of (EF), is expressed as hexInEF in EF path filter bandwidth range.The case where the third fluorophor is not present
Under, the contribution ratio of FAM fluorophor is famInAB/ (famInAB+hexInAB), HEX fluorescent base in the channel AB in the channel AB
The contribution ratio of group is hexInAB/ (famInAB+hexInAB)；The contribution ratio of FAM fluorophor is in the channel EF
FamInEF/ (famInEF+hexInEF), the contribution ratio of HEX fluorophor is hexInEF/ (famInEF+ in the channel EF
hexInEF)。
Similarly, in embodiments of the present invention, it by taking three kinds of fluorophors as an example, and is said by taking first passage unit as an example
Bright, the luminous contribution ratio of each fluorophor is obtained by following steps：
The luminous spectrogram of the corresponding calibration of three kinds of fluorophors is calculated in first passage unit (corresponding filtering bandwidth range)
Interior integral, using the luminous contribution as corresponding fluorophor in the channel unit；
By luminous contribution of first fluorophor in first passage unit divided by three kinds of fluorescence in first passage unit
The luminous contribution summation of group obtains first fluorophor in the luminous contribution ratio a1 of first passage unit1, similarly
To the second fluorophor first passage unit luminous contribution ratio a21With third fluorophor first passage unit hair
Light contribution ratio a31.The first fluorophor is similarly obtained again in the luminous contribution ratio a1 of second channel unit2, the second fluorescent base
Luminous contribution ratio a2 of the group in second channel unit2With third fluorophor second channel unit luminous contribution ratio
a32；And first fluorophor third channel unit luminous contribution ratio a13, the second fluorophor is in third channel unit
Luminous contribution ratio a23With third fluorophor third channel unit luminous contribution ratio a33。
S3, the luminous contribution ratio according to each fluorophor in the luminous intensity of the channel unit and the channel unit
Rate obtains the concentration of each fluorophor.
Specifically in the embodiment of three fluorophors of the invention, simultaneous following equation：
INTENSITY1=a11×c1+a21×c2+a31×c3；
INTENSITY2=a12×c1+a22×c2+a32×c3；
INTENSITY3=a13×c1+a23×c2+a33×c3；Wherein, c to be solved1For the hair of first fluorophor
Luminous intensity, c2For the luminous intensity of second fluorophor, c3For the luminous intensity of third fluorophor, INTENSITY1、
INTENSITY2And INTENSITY3For the luminous intensity for detecting obtained each channel, a11、a21、a31、a12、a22、a32、
a13、a23And a33It has been acquired in above-mentioned S2, solve system of equation obtains c1、c2And c3, i.e., the luminous intensity of each fluorophor.
It should be noted that method is same when target spot quantity (i.e. the number of species of fluorophor) are more than two or three
The concentration measuring method of three kinds of fluorophors obtains the luminous intensity of each fluorophor by following formula：
INTENSITYi=a1i×c1+a2i×c2+......+aji×cj, wherein target spot quantity and for indicating target spot
Fluorophor number of species be j, i is channel number, INTENSITYiFor the strong light for detecting i-th obtained of channel
Degree, c1For the luminous intensity of first fluorophor, c2For the luminous intensity of second fluorophor, cjFor j-th of fluorophor
Luminous intensity, a1iFor the luminous contribution ratio of first fluorophor in i-th of channel, a2iIt is in i-th of channel
The luminous contribution ratio of two fluorophors, ajiFor the luminous contribution ratio of j-th of fluorophor in i-th of channel, 2≤i
≤j.Another distinctive points of the present invention and the prior art are that in the prior art fluorescence crosstalk in order to prevent, glimmering in two kinds of detection
It there is certain error when light radical concentration, when detecting three kinds of fluorophors, biggish error can be generated, in the prior art
Three kinds or more of fluorophor will not be substantially measured with fluorescent marker probe method, and the measurement method of the application reacts PCR
There is no limit can accurately calculate the light intensity of each fluorophor by accurately calculating to target spot quantity in sample.Pass through this
After the PCR algorithm that invention provides is corrected, " the grey point " observed in different channels can be effectively reduced.
And further include after the luminous intensity for obtaining each fluorophor：According to the concentration of pre-established fluorophor with
The relational model of luminous intensity obtains the corresponding concentration of luminous intensity of the fluorophor.For example, according to the glimmering of known concentration
Light group measures its light intensity, concentration/the Relationship of Light intensity model (concentration/light intensity curve) of fluorophor is established, according to the relationship mould
The luminous intensity of obtained fluorophor is converted corresponding concentration by type.The concentration of target spot be by count it is all negative and
The ratio of positive drop calculates.And our algorithm is the radical concentration calculated in some specific drop, this concentration
Calculating facilitates us and judges that this drop is negative or positive.The application by accurately calculate drop in fluorophor it is dense
Degree judges whether some fluorophor in drop is negative or the positive according to radical concentration, this passes through light with traditional method
Judge that the Yin/Yang of drop in some channel is compared by force, the measurement result of the application has more reliability.
In another embodiment of the present invention, a kind of droplet type digital pcr system is provided, using such as above-described embodiment
The measurement method is realized to each droplet quantitative observation.
To sum up, the digital pcr of measurement of concetration is carried out the present invention provides a kind of PCR density calculating method and using the algorithm
System is corrected by algorithm, " the grey point " observed in channel is effectively reduced, and correct interchannel brought by multiple target spot
Fluorescence crosstalk；Except this can not limit the bandwidth of filtering channel, specially using the calculation method of the application so as to improve
The port number of Multiple detection.
The foregoing is merely presently preferred embodiments of the present invention, is not intended to limit the invention, it is all in spirit of the invention and
Within principle, any modification, equivalent replacement, improvement and so on be should all be included in the protection scope of the present invention.
Claims (10)
1. a kind of multiple digital pcr concentration measuring method, which is characterized in that be suitable for while detecting in a PCR response sample
Two or more target spots concentration, different target spots indicated using different fluorophor, the concentration measuring method
Including：
Each corresponding fluorophor, determines filtering bandwidth range, and with the corresponding channel unit of determination, and it is single to detect each channel
The luminous intensity of member；
According to the luminous spectrogram of fluorophor filtering bandwidth range corresponding with the fluorophor, obtain each in channel unit
The luminous contribution ratio of fluorophor；
According to the luminous contribution ratio of each fluorophor in the luminous intensity of the channel unit and the channel unit, obtain
The concentration of each fluorophor.
2. multiple digital pcr concentration measuring method according to claim 1, which is characterized in that described according to the channel
The luminous contribution ratio of each fluorophor in the luminous intensity of unit and the channel unit, obtains the dense of each fluorophor
Degree includes：
The luminous intensity of each fluorophor is obtained by following formula：
INTENSITYi=a1i×c1+a2i×c2+......+aji×cj, wherein target spot quantity and the fluorescence for indicating target spot
The number of species of group are j, and i is channel number, INTENSITYiFor the luminous intensity for detecting i-th obtained of channel, c1It is
The luminous intensity of one fluorophor, c2For the luminous intensity of second fluorophor, cjFor the strong light of j-th of fluorophor
Degree, a1iFor the luminous contribution ratio of first fluorophor in i-th of channel, a2iFor second fluorescence in i-th of channel
The luminous contribution ratio of group, ajiFor the luminous contribution ratio of j-th of fluorophor in i-th of channel, 2≤i≤j.
3. multiple digital pcr concentration measuring method according to claim 2, which is characterized in that obtaining each fluorescent base
Further include after the luminous intensity of group：
According to the relational model of the concentration of pre-established fluorophor and luminous intensity, the luminous intensity of the fluorophor is obtained
Corresponding concentration.
4. multiple digital pcr concentration measuring method according to claim 1, which is characterized in that determined to fluorophor
Further include before filtering bandwidth range：Selection is for the corresponding fluorophor for indicating different target spots, the wave of multiple fluorophors
Long range registration is greater than the corresponding wavelength difference of emission peak of preset first threshold and/or the multiple fluorophor
Greater than preset second threshold.
5. multiple digital pcr concentration measuring method according to claim 1, which is characterized in that for indicating different target spots
Fluorophor include but is not limited to FAM, TET, JOE, HEX, CY3, TAMRA, ROX, Texas Red, LC RED640 and other
Fluorophor, the corresponding calibration of each fluorophor have luminous spectrogram.
6. multiple digital pcr concentration measuring method according to claim 1, which is characterized in that in the channel unit
The luminous contribution ratio of each fluorophor is obtained by following steps：
Calculate different fluorophors shine spectrogram the corresponding filtering bandwidth range of the same channel unit integral, using as right
Answer luminous contribution of the fluorophor in the channel unit；
By luminous contribution of one of fluorophor in the channel unit divided by fluorophors all in the channel unit
Luminous contribution summation, obtain the fluorophor in the luminous contribution ratio of the channel unit.
7. multiple digital pcr concentration measuring method according to claim 1, which is characterized in that each is glimmering for the correspondence
Light group determines that filtering bandwidth range includes：
According to the wavelength characteristic of each fluorophor, the filter of corresponding selection different frequency bands width, the filter
Frequency bandwidth is covered and is uniquely covered in the emission peak of corresponding fluorophor and/or the frequency bandwidth of the filter
The coincidence wave-length coverage of multiple fluorophors is greater than preset third threshold value；
Bandwidth with the frequency bandwidth of the filter of selection, as the corresponding channel unit of fluorophor.
8. multiple digital pcr concentration measuring method according to claim 7, which is characterized in that each channel of detection
The luminous intensity of unit includes：
PCR response sample is filtered using selected filter one by one respectively；
Light-intensity test is carried out to the PCR response sample being filtered, obtains the luminous intensity of corresponding channel unit one by one.
9. multiple digital pcr concentration measuring method according to claim 1, which is characterized in that in the PCR response sample
Target spot quantity be two, three or three or more.
10. a kind of droplet type digital pcr system, which is characterized in that using the measurement as described in any one of claim 1-9
Method is realized to each droplet quantitative observation.
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| CN201810642098.9A CN108841939B (en) | 2018-06-21 | 2018-06-21 | Multi-digital PCR concentration measuring method and micro-drop type digital PCR system |
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Denomination of invention: Multiple digital PCR concentration measurement method and droplet digital PCR system Effective date of registration: 20230726 Granted publication date: 20200922 Pledgee: Fengxian Branch of Shanghai Rural Commercial Bank Co.,Ltd. Pledgor: BEIJING ZHIYU BIOTECHNOLOGY Ltd. Registration number: Y2023310000403 |