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Method for building buckwheat AFLP reaction system and application of buckwheat AFLP reaction system

A technology of reaction system and method establishment, applied in the field of biology

Inactive Publication Date: 2014-01-01
SHENZHEN SUNSMILE BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The use of AFLP technology has been widely used in the research of rice, corn, soybean, wheat, millet and other crops at home and abroad, but there are few reports on the application of buckwheat

Method used

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  • Method for building buckwheat AFLP reaction system and application of buckwheat AFLP reaction system
  • Method for building buckwheat AFLP reaction system and application of buckwheat AFLP reaction system
  • Method for building buckwheat AFLP reaction system and application of buckwheat AFLP reaction system

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] The establishment method of the AFLP reaction system of buckwheat, see figure 1 shown, including the following steps:

[0056] S11, preparation of template DNA: Weigh 0.5 g of fresh leaves of Shaanxi Baizhong buckwheat from Inner Mongolia, put them in a mortar, pass through liquid nitrogen to pre-cool, then put the leaves in liquid nitrogen and grind them evenly until they are all ground to powder ; Then add 600ul of CTAB extract and react at a constant temperature of 65°C for 1 hour, wherein the CTAB extract contains 2.5% cetyltrimethylammonium bromide (CTAB), 150mmol / L Tris-HCl, 30mmol / L of ethylenediaminetetraacetic acid (EDTA), 1.6mol / L of NaCl, 1% of β-mercaptoethanol, and 2% of polyvinylpyrrolidone (PVP); then cool to room temperature, add the above-mentioned phenol-chloroform solution to extract 2 times to obtain the template DNA; the quality of the extracted template genomic DNA was detected by 1% agarose gel electrophoresis, and stored at -20°C for later use;...

Embodiment 2

[0062] The establishment method of the AFLP reaction system of buckwheat, see figure 1 shown, including the following steps:

[0063] S21, preparing template DNA: Weigh 0.5 g of fresh leaves of Lingqiu tartary buckwheat produced in Shanxi, place them in a mortar, pass through liquid nitrogen to pre-cool, then place the leaves in liquid nitrogen and grind them evenly until they are all ground to powder; then Add 550ul of CTAB extract and react at a constant temperature of 65°C for 1 hour, wherein the CTAB extract contains 2% cetyltrimethylammonium bromide (CTAB), 150mmol / L Tris-HCl, 20mmol / L ethylenediaminetetraacetic acid (EDTA), 1.6mol / L NaCl, 1% β-mercaptoethanol, 1% polyvinylpyrrolidone (PVP); then cooled to room temperature, adding the above phenol-chloroform solution for extraction 3 times , to obtain template DNA; the quality of the extracted template genomic DNA was detected by 1% agarose gel electrophoresis, and stored at -20°C for future use;

[0064] S22, Digest te...

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Abstract

The invention provides a method for building a buckwheat AFLP reaction system. According to the method for building the buckwheat AFLP reaction system, reaction conditions of AFLP critical steps are improved, and the number of extraction times is increased properly, so that not only are impurities in reaction liquid removed fully, but also mechanical breakage of buckwheat DNA is reduced and quality of extraction of the DNA is improved; digestion is fully performed on buckwheat genome DNA for the set digestion time; the set Mg2+ concentration can improve the output rate of product specific DNA and avoid amplification of non-specific DNA, the reaction system which is good in reliability, strong in repeatability, high in credibility, convenient and quick to achieve and suitable for buckwheat is built, few DNA samples are needed, Southern hybridization is not needed, and serial information of the DNA does not need to be known in advance. The method is simple in process and easy to operate, the output amount is high, and production cost is lowered.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for establishing a buckwheat AFLP reaction system and its application. Background technique [0002] Buckwheat is a kind of economic crop with both food and medicine. It is rich in nutrients, rich in protein, fatty acid, vitamins and minerals, etc. Buckwheat protein has high biological potency, and its amino acid composition is reasonable. Because of the high content of arginine, lysine, tryptophan and histidine, buckwheat has a good complementarity with other cereal grains. Buckwheat is not only nutritious, but also rich in bioflavonoids, polypeptides, sugar alcohols and D-chiro-inositol and other highly active medicinal ingredients, which have the functions of lowering blood sugar, fat, cholesterol, anti-oxidation, anti-aging and scavenging free radicals , the bioactive components contained in buckwheat have attracted widespread attention from the medical comm...

Claims

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Application Information

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IPC IPC(8): C12N15/10C12Q1/68
Inventor 潘韵侯雅君李永新郑文官陈义昆唐明辉
Owner SHENZHEN SUNSMILE BIOTECH
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