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Preservative and preservation method for intermediate PCR (Polymerase Chain Reaction) sequencing product

A preservation method and preservative technology, applied in biochemical equipment and methods, microbial measurement/inspection, etc., can solve the problems of inconvenient arrangement, expensive, and gel consumption of experimental operations, and achieve the effect of improving the success rate of sequencing on the machine

Active Publication Date: 2014-09-10
CHENGDU ZHONGCHUANG QINGKE MEDICAL LAB CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The gels used in sequencing are generally imported reagents, which are relatively expensive
For small laboratories or hospitals, the sample size for sequencing is not large, and it takes a lot of gels to sequence a few sites. At the same time, the experimental operation is very inconvenient to arrange, and several samples must be completed in a short time.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0012] Preservative preparation: Take sodium acetate, disodium edetate and glycogen, add solvent and mix well, adjust pH, the solvent can be water or ethanol, etc. The final concentration of mixed sodium acetate was 1 mol / L, the concentration of disodium edetate was 0.03 mol / L, and the concentration of glycogen was 3 g / L.

[0013] The PCR sequencing intermediate product may be a product generated after the sequencing PCR step or the sequencing PCR product purification step using Sanger sequencing method. Add a preservative that is a quarter of the volume of the PCR sequencing intermediate product into the PCR sequencing intermediate product, mix well, and store at minus 20°C.

Embodiment 2

[0015] Preservative preparation: Take sodium acetate, disodium edetate and glycogen, add solvent and mix well, adjust pH, the solvent can be water or ethanol, etc. The final concentration of sodium acetate after mixing was 1.2 mol / L, the concentration of disodium edetate was 0.04 mol / L, and the concentration of glycogen was 4 g / L.

[0016] The PCR sequencing intermediate product may be a product generated after the sequencing PCR step or the sequencing PCR product purification step using Sanger sequencing method. Add a preservative that is a quarter of the volume of the PCR sequencing intermediate product into the PCR sequencing intermediate product, mix well, and store at minus 20°C.

Embodiment 3

[0018] Preservative preparation: Take sodium acetate, disodium edetate and glycogen, add solvent and mix well, adjust pH, the solvent can be water or ethanol, etc. The final concentration of sodium acetate after mixing is 1.5 mol / L, the concentration of disodium edetate is 0.05 mol / L, and the concentration of glycogen is 5 g / L.

[0019] The PCR sequencing intermediate product may be a product generated after the sequencing PCR step or the sequencing PCR product purification step using Sanger sequencing method. Add a preservative that is a quarter of the volume of the PCR sequencing intermediate product into the PCR sequencing intermediate product, mix well, and store at minus 20°C.

[0020] control group

[0021] Add a solvent that is a quarter of the volume of the PCR sequencing intermediate product to the PCR sequencing intermediate product, mix well and store at minus 20°C.

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PUM

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Abstract

The invention discloses a preservative for an intermediate PCR (Polymerase Chain Reaction) sequencing product. The preservative comprises 1-1.5mol / L sodium acetate, 0.03-0.05mol / L ethylene diamine tetraacetic acid disodium salt and 3-5g / L glycogen. The preservative disclosed by the invention can be used for increasing the success rate of computer sequencing of the preserved intermediate PCR sequencing product.

Description

technical field [0001] The invention relates to the preservation of biological preparations, in particular to a preservative for intermediate products of PCR sequencing and a preservation method using the preservative. Background technique [0002] DNA is an important component of chromosomes. It is the genetic material of eukaryotes and contains all the genetic information of individual species. It can be used to construct genome libraries, isolate required genes, and detect related molecular markers. DNA is often amplified using a PCR amplifier and sequenced using Sanger sequencing. [0003] Sanger sequencing is currently a widely used direct sequencing method. Whether it is a large hospital, a laboratory, or a small research institution, it is possible to use Sanger sequencing for detection or research. This method is low-cost, fast, accurate, etc. features. The sequencing steps of the Sanger sequencing method can be roughly divided into five steps: template PCR, templa...

Claims

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Application Information

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IPC IPC(8): C12Q1/68
CPCC12Q1/6806C12Q2527/125
Inventor 石昌木李晓瑞朱智鹏刘艺张蓉
Owner CHENGDU ZHONGCHUANG QINGKE MEDICAL LAB CO LTD