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Rapid soybean DNA obtaining method for PCR amplification and PCR amplification method

A soybean, fast technology, applied in biochemical equipment and methods, microbial determination/inspection, etc., can solve the problems of high cost, many operation steps, irritating taste in the extraction process, etc., achieving low cost, high sample loading efficiency, and operation. simple effect

Inactive Publication Date: 2020-07-28
RICE RES ISTITUTE ANHUI ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The extraction steps are complicated, there are many types of chemical reagents prepared, the extraction process has a strong irritating smell, and the operation steps are many or the cost is high, which is not conducive to high-throughput molecular assisted breeding (see Table 1).

Method used

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  • Rapid soybean DNA obtaining method for PCR amplification and PCR amplification method
  • Rapid soybean DNA obtaining method for PCR amplification and PCR amplification method
  • Rapid soybean DNA obtaining method for PCR amplification and PCR amplification method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1: Extraction of soybean DNA

[0028] 1. Material handling

[0029] Take 1 cm of petiole of a certain soybean variety, cut it into pieces, and set aside;

[0030] 2. Reagent preparation

[0031] Prepare reagent A, which is a NaOH solution with a concentration of 4 g / L;

[0032] Reagent B is prepared, and the formula for each 500mL of Reagent B is as follows: 1mol / L Tris-HCl 5ml with pH 8.0, 1ml EDTA 0.5mol / L, add distilled water to make up to 500ml;

[0033] 3. Extraction of soybean DNA

[0034] Add the chopped petiole tissue in step 1 to 60ml of reagent A, place it in a PCR machine, cook at 95-100°C for 3 minutes, then add 60ml of reagent B, stir evenly, and then the DNA template of soybean tissue can be obtained.

Embodiment 2

[0036] The extraction process of this example is the same as that of Example 1, the difference is that in step 3, it is boiled for 6 minutes at 95°C.

Embodiment 3

[0038] The extraction process of this example is the same as that of Example 1, the difference is that in step 3, it is boiled at 97° C. for 9 minutes.

[0039] After testing, among Examples 1-3, the DNA yield rate of Example 1 was the highest, which was 100%, and the time was the shortest. It was determined that Example 1 was the optimal solution.

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Abstract

The invention discloses a rapid soybean DNA obtaining method for PCR amplification and a PCR amplification method. Alkali-treated soybean petioles are directly used as a PCR reaction template, and plant DNA can be rapidly, effectively, simply and conveniently extracted to obtain a soybean tissue DNA template; and PCR amplification is carried out through the DNA template by using an SSR standard primer. The method has the technical effects of high timeliness, simple extraction steps, less chemical reagent, small smell and environmental protection when being used for seed nature detection and molecular marker assisted breeding.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for rapidly obtaining soybean DNA for PCR amplification and a PCR amplification method. Background technique [0002] With the development of molecular biology technology, molecular marker technology based on PCR (polymerase chain reaction) has been widely used in the analysis of genomic DNA because of its relative simplicity and speed, thus greatly promoting genetic breeding and genetic evolution. , Germplasm classification and research and development of gene cloning, gene location and so on. With the application of various PCR-based molecular markers, the acquisition of DNA templates is the basis of all this. According to the research object, research purpose and research cost of plants, the methods used to extract genomic DNA are also different. Among them, with the wide application and promotion of molecular-assisted crop breeding, genotype identification ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6806
CPCC12Q1/6806C12Q2531/113C12Q2565/125
Inventor 许学汪秀峰马卉鲍翔宇吴爽刘文博左泽平麦霄黎潘伟芹林玲
Owner RICE RES ISTITUTE ANHUI ACAD OF AGRI SCI
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