CN102646170A - DNA (Deoxyribo Nucleic Acid), RNA (Ribo Nucleic Acid) and albumen glue electronization standard and analysis application software - Google Patents

Abstract

The invention provides a DNA, RNA and albumen glue electronization standard and analysis application software. Currently, people spend a great amount of research money in a scientific research due to a non-reversible property of reference substance of DNA (Deoxyribo Nucleic Acid), RNA (Ribo Nucleic Acid) and albumen glue in use; and the DNA, the RNA and the albumen glue may cause serious biological pollution to the environment. The invention provides the DNA, RNA and albumen glue electronization standard and analysis application software, wherein the DNA, the RNA and the albumen glue have stable mobility at experiment conditions of certain time, temperature, glue concentration, voltage and current on electrophoresis apparatuses with a uniform specification, DNA, RNA and protein biological reference substance actually applied in the DNA, so that the RNA and the albumen glue can be replaced by building a computer software database, and designing and plaiting the standard application software through a great amount of experiment data. Verified by repeatedly testing, the accuracy of the product reaches 99.90-100%. The product can be repeated, has strong stability and can be used for entirely replacing the biological reference substances actually added in the DNA, the RNA and the albumen glue. The product has important meanings on saving research cost, reducing biological pollution to the environment, and providing stable electronic standard in the DNA, the RNA and the albumen glue, and great social benefits can be realized.

Description

DNA, RNA and protein adhesive electronization standard and analytical applications software

Technical field this invention belongs to the molecular biotechnology application; He can be applicable to all biologies (are comprised animal; Plant, microorganism etc.) DNA, the testing process of RNA and protein example; For these sample detection provide DNA, RNA and protein electronization standard and analytical applications software.

Background technology is in molecular biology experiment, and the researchist is dna fragmentation (restriction enzyme section section in the detection of biological sample; Pcr amplified fragment; During dna fragmentations such as molecular cloning detect); When inspection detects gene expression product mRNA; In detecting the protein level variation, all to carry out the separation of agarose gel earlier, dyeing and evaluation.Wherein, In the actual biological sample detachment process of carrying out each piece glue, all want to add known DNA at first swimming lane simultaneously; RNA and protein biological standard sample; With its DNA known in glue, RNA clip size, protein molecular weight difference are used as fragment and the molecular weight size that the detected object in the test sample is wanted in the canonical reference prompting.So, DNA, RNA and protein standard model are at DNA, requisite biological products in RNA and the protein biological sample isolation identification.But because of it makes complicated; Cost is high and possibly after usefulness, can bring biological pollution to environment; So set up DNA, it will be the significant change in the biological sample qualification process that RNA and protein electronic standard figure replace traditional biological products, and it will produce huge economic and social benefit.

DNA, in addition purifying is expressed behind the protein of fixed dna fragment and fixed member amount of known gene also through molecular cloning by used biological standard article system on RNA and the protein adhesive electrophoresis, combines.Its complex manufacturing technology, the cycle is long, and cost is high.And with after can't recycling, have potential biological pollution dangerous.

The present invention adopts the digital collection of biotic district band, sets up data bank and computer software composite drawing, can replace DNA fully; The biological standard produce article that RNA and albumen are traditional; Be to practice thrift living resources, environment purification provides the DNA of accurate practicality; RNA, albumen electronization standard is laid a good foundation.

Traditional DNA, the method for production of RNA and protein biology standard model:

1. traditional DNA biological standard sample is that the dna fragmentation with designated length is cloned in the plasmid vector; Be transfected into bacterium then; Extract DNA behind the massive duplication; Cut out dna fragmentation and carry out the large-scale purification separation with restriction enzyme, mix in proportion with the DNA of different fragments length at last, be mixed with DNA biological standard sample.

2.RNA the biological standard sample is the RNA separation and purification that regular length is expressed, the RNA fragment with other different lengths is mixed into RNA biological standard sample then.

3. the protein biology standard model is that known molecular size (molecular weight) expressed proteins is cloned; Carry out transfection or vivoexpression then; Expressed proteins is carried out separation and purification, and the protein purification thing of last different molecular weight mixes the back in proportion and forms the protein biology reference material.

SUMMARY OF THE INVENTION Core of the present invention include: First, the use of bio-products company acquired DNA, RNA and protein biological standard samples, using its standard electrophoresis tank systems, at a fixed concentration of glue, fixed time, a fixed voltage, shed (for example: 1% gel at 80 volts, go gum solution in 1xTAE 30 minutes) for separation gel, and ethidium bromide (disseminated DNA, RNA gel) chloride Silver (stained protein gel) after staining after separation of the standard zone and measuring the distance between the origin of the data collection and the formation of databases, preparation of gel image analysis computer software (software with mining plan, the establishment of a database, build a new figure to fight the formation of new icon computing platform, set the analysis, calculation function, reserve function, transfer function) on which the application software into electrical standard zone map, this map Electronics Standards zone with actual specimens software to obtain DNA, RNA, and protein gel separation image (with a ruler calibrated image) and then spliced into new images (including electronic standard and gel separation zone of the actual sample zone), for subsequent analysis, storage and use.

Electronization DNA, RNA, the manufacturing process of protein standard and application software and main points:

1. adopt the biological DNA that buys with flat company, RNA and protein biology reference material use it in standard electrophoresis tank system, in fixing proud concentration; Regular time, carry out glue under the fixing voltage, electric current and separate, use the pyridine of bromine second (to infect DNA then; RNA glue) separate the measurement of distance between laggard Xingqi and the last appearance initial point with the laggard column criterion of silver chloride (dying protein adhesive) dyeing district band, data are compiled and are formed data bank, and (software possesses the figure of adopting to establishment computing machine glue imaging analysis software, sets up database; Assembly is built new figure and is formed new diagram computing platform, analysis is set, computing function; Reserve function, transmitting function), in its application software, data information is changed into electronic standard district band figure (see figure 1); What each electronic standard district band figure showed is at fixing glue concentration, voltage, electric current; Point sample initial point and each standard regions band under the fixing glue disengaging time condition (yardstick that contains demarcation: going up sampling point is zero, and the distance of standard regions band that on glue, separates and initial point is demarcated), the available fixing glue concentration of each electronic standard district band figure; Voltage, electric current, fixing glue disengaging time are retrieved from the software document picture library and are taken out.

2. computer process (contain and adopt figure, set up database, piece together and to build new figure and form new diagram computing platform, analysis is set, computing function, reserve function, transmitting function).Utilize ripe at present computer software establishment technique and means, establishment possesses the figure of adopting, and sets up database; Form electronic standard district band figure, assembly is built new figure and is formed new diagram computing platform, and analysis is set; Computing function; Reserve function, the DNA of transmitting function, RNA and imaging of protein electronic standard glue and analyzing software system.The basic function of these software systems and technical support condition are described below:

Multiple figure obtains form and supports standard, and the twain interface can obtain image from the industrial standard input equipment.The video input apparatus of support standard usb interface.Support standard directshow (wdn) video obtains technology.

Abundant image processing capacity image processing function: contrast; Hsv, dhsl; Rgb; Sharpening; Fft; Binary conversion treatment, gray scale processing etc.Descriptive geometry is handled: left rotation and right rotation; The horizontal vertical rotation; By the angle rotation; Control rotation by hand; Cut out by hand etc.Function internal memory recovery technology is efficiently added in image operation control design, caption text.

Powerful experimental analysis functional support is carried out the expert data analysis to electrophoretic image, can accurately test numerous test datas such as brightness values, sample area, OD value, average light ratio, square light ratio value in each constituency.Selecting of the manual rectangle of system's support, circle and oval constituency; And can add the mark numbering automatically to each constituency; Setpoint color and font size etc. are supported the input of accurate data locking in constituency and data size simultaneously, improve the position and the dimensional accuracy in constituency.Support the duplicating of constituency, stickup and deletion action, support manual selection a plurality of constituencies, and the unified integer function of the size that can carry out all constituencies.Convenient, flexible benchmark constituency comparison function can arbitrarily specify certain constituency to compare the constituency as benchmark, analyses and compares with the benchmark constituency in other constituencies.Specialty is controlled design easily, when being supported in the constituency and setting, uses mouse that the constituency is pulled and adjusts, and simultaneously, the mouse scaling operation to base map is provided also, makes man-machine interaction and convenient and efficient.The duty hold function of providing convenience can save as item file with current working state, and supports the strange land of item file to open and handle, and improves the convenience of research work, has reduced the passivity of research work.Simultaneously, also can carry out confidential treatment, promote the security requirement of test figure greatly item file.Adopt computer graphic image Processing Algorithm efficiently, greatly improved processing speed, and support WYSIWYG (What You See Is What You Get) pattern, all operations generates treatment effect immediately.

The image output machine printing function of specialty supports the direct preservation of image and format conversion to preserve, and algorithm is efficiently quick.Image is directly printed in supporting to handle, and supports Print Preview, and has the function of setting type easily.Support the printing and the preservation of specialty analysis report, support Print Preview, and the importing and the printing of historical analysis report.

Powerful analysis result information functional analysis result provides with the expert data forms mode, can convenient printing preview and preservation, and can repeatedly open the print file of preserving, print output.During Print Preview, can carry out zoom operations, and support the test picture after handling is printed the demonstration of report.System supports printed report is exported to html format and MS EXCEL form simultaneously, supports the synchronous export function of test picture.During the reporting printing of system, provide standard print that function is set, convenient various printers and the input equipment of connecting.

System configuration requires basic configuration: operating system: MS Windows XP series, Vista series, Windows7 series.CPU:Intel P4, Intel match poplar series etc.Internal memory: 1G comprises operating system and database required memory.Hard disk: 12M (software program, printer etc.).Standard device: keyboard, mouse, printer etc.Other: test is obtained equipment etc. with video image.

Various gel type such as intelligent gel electrophoresis photo comparison functional support standard DNA, RNA and protein adhesive, and the choosing of underlying parameters such as corresponding glue voltage, gum concentration and glue duration, and can generate the gel electrophoresis picture of standard automatically according to selected parameter.Support merges to the gel electrophoresis picture that generates on the test picture, convenient compare of analysis to experimental data.Support to the gel electrophoresis picture that generates be rotated operation, stack operation, zoom operations, drag operation etc., support is carried out the adjustment of size dimension to it.Aggregate speed is fast, merges when supporting a plurality of master sample and handles.According to the technical parameter of different displays, generate the standard scale automatically, make things convenient for actual measurement.

3. digitizing DNA, the basic functional principle of RNA and protein standard and analytical applications software:

Carry out DNA by defined terms, (gum concentration is walked the glue time, voltage in the glue separation of RNA and protein sample; Electric current), through the pyridine of bromine second (DNA, RNA), silver chloride (albumen) dyes behind the glue with transparent plastic footage number Ding Caitu; Then with software in the electronic standard figure of the same terms splice (use fixing glue concentration, voltage, electric current, fixing glue disengaging time take out electronic standard figure retrieval) from the software document picture library; Form new figure, can analyze, calculate, store.

Its concrete operations step is following:

1. the gum concentration of configuring fixed.Prepare to want DNA, RNA and the protein sample of separation detection.

2. in electrophoresis tank, press fixing voltage, electric current is walked the glue of regular time length.

3. (DNA, RNA), silver chloride (albumen) dyes glue in bromine second pyridine.

4., the transparent plastic ruler adopts figure under demarcating

5. the DNA that gathers; (ruler that contains demarcation: going up sampling point is zero for RNA or protein sample glue separation graph; The distance of district's band that on glue, separates and initial point is demarcated) with software in the electronic standard figure (yardstick that contains demarcation: going up sampling point is zero, the distance quilt demarcation of the standard regions band that on glue, separates and initial point) of the same terms splice.

6. cutting assembles new figure.

7. analyze, store, transmit.

Claims (5)

1. this patent has proposed DNA in the world first, the notion of RNA and protein adhesive standard regions band electronization, and made the electronic standard application software first, its core technology is to utilize DNA; RNA fragment protein biological standard district band is at fixing gum concentration, voltage, and its stable mobility has collected reliable data during current constant; Data are compiled and are formed data bank, and (software possesses the figure of adopting to establishment computing machine glue imaging analysis software, sets up database; Assembly is built new figure and is formed new diagram computing platform, analysis is set, computing function; Reserve function, transmitting function), in its application software, data information is changed into electronic standard district band figure; What each electronic standard district band figure showed is at fixing glue concentration, voltage, electric current; Point sample initial point and each standard regions band under the fixing glue disengaging time condition (yardstick that contains demarcation: going up sampling point is zero, and the distance of standard regions band that on glue, separates and initial point is demarcated), the available fixing glue concentration of each electronic standard district band figure; Voltage, electric current, fixing glue disengaging time are retrieved from the software document picture library and are taken out.In experiment; The DNA that gathers; (ruler that contains demarcation: going up sampling point is zero for RNA or protein sample glue separation graph; The distance of district's band that on glue, separates and initial point is demarcated) with software in the electronic area band standard drawing (yardstick that contains demarcation: going up sampling point is zero, the distance quilt demarcation of the standard regions band that on glue, separates and initial point) of the same terms splice.Cutting assembles new figure.Analyze, store, transmit.The application's request is to DNA, the notion of RNA and protein adhesive standard regions band electronization, and electronic standard application software and manufacturing technology thereof, program and follow-up modification propose patent protection.

2. the DNA biological standard district band of the different fragments length that proposes first is converted into the notion and the method for making of electronic standard regions band.

3. the RNA biological standard district band of the different fragments length that proposes first is converted into the notion and the method for making of electronic standard regions band.

4. the protein biological standard district band of the different molecular weight that proposes first is converted into the notion and the method for making of electronic standard regions band.

5. electronic standard application software and manufacturing technology thereof, program and the follow-up modification made first.