Extraction method of wheat genome DNA

An extraction method and genome technology are applied in the field of wheat genome DNA extraction, which can solve the problems of cumbersome and time-consuming extraction steps, poor DNA extraction quality, and complicated operations, simplifying the steps of DNA extraction, and improving the efficiency of sample addition and extraction. The effect of work efficiency

Inactive Publication Date: 2015-06-10
NORTHWEST A & F UNIV
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Problems solved by technology

However, the traditional CTAB method is cumbersome to operate, consumes a lot of reagents, and uses many utensils. The grinding utensils need to be reused, which is easy to cause sample contamination and low yield.
[0003] At present, most of the reports on plant DNA extraction methods are improved on the basis of SDS method and CTAB method, but none of them can get rid of the operation steps of grinding samples one by one with grinding rods or glass rods, and the extraction steps are cumbersome and t...

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  • Extraction method of wheat genome DNA
  • Extraction method of wheat genome DNA
  • Extraction method of wheat genome DNA

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Embodiment 1

[0027] 1. DNA extraction

[0028] (1) Sow and germinate the seeds of the two F2 generation populations. After the seedlings are 5 cm high, it generally takes 5 to 7 days. After the first true leaf is unfolded, take about 5 cm of young wheat leaves (dry weight is about 50 mg), and divide it into Several sections are put into eight-tube tubes in corresponding order (the capacity of each test tube is 1ml), and each 96-well test tube box is equipped with 12 rows of eight-tube tubes.

[0029] (2) After sampling, use a bead setter to put a steel ball (diameter 4mm) that has been cleaned with absolute alcohol in each test tube, then open the sample and place it in a vacuum freeze dryer, and dry it at -110°C for 24 hours .

[0030] (3) Take out the test tube box, quickly cover the tube cover and the grinding sample box cover, fix the test tube box on the grinder, pay attention to keep the balance on both sides, and grind, generally 25-30 times / second, about 1 minute, and show the sam...

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Abstract

The invention discloses an extraction method of a wheat genome DNA. The method comprises the following steps: putting wheat material samples to be extracted into 96-well plates respectively; then putting steel balls into each hole; uncovering and putting into a vacuum freezing dryer to be subjected to freeze drying for 20-28 hours at the temperature of -100-(-120) DEG C; grinding the wheat material samples by using a grinding instrument; finally, extracting DNA from the grinded powder samples by using a discharge gun in an improved SDS method. According to the extraction method, wheat tissue vanes are pulverized by using the grinding instrument after vacuum freeze drying is carried out; compared with a traditional liquid nitrogen grinding method, the extraction method has the advantages that the labor force is greatly released, and the working efficiency is improved; meanwhile, by using 96-well plates and the discharge gun, the sample extraction efficiency is greatly improved. The extraction method of the wheat genome DNA is convenient, fast, high in flux and high in quality.

Description

technical field [0001] The invention relates to a plant genome DNA extraction method, in particular to a wheat genome DNA extraction method. Background technique [0002] With the continuous development of molecular biology, the demand for samples in many molecular biology experiments is very large. Simple, fast, efficient and stable DNA extraction technology is a necessary prerequisite for promoting the development of wheat-related research at the molecular level, and it is also the key to research efficiency. The DNA extraction of the wheat genome generally needs to go through several steps such as breaking the cell wall, lysing the cell membrane, removing secondary substances such as polysaccharides and polyphenols, denaturing and separating proteins, precipitating nucleic acids, removing RNA, and concentrating DNA. Generally speaking, the CTAB (cetyltriethylammonium bromide) method has unique advantages in processing plant tissues containing polysaccharide components, an...

Claims

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Application Information

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IPC IPC(8): C12N15/10
Inventor 吴建辉王琪琳韩德俊康振生曾庆东穆京妹薛文波许鑫徐琪沈川吕兴娜王浩庭王美杰李海洋苏萍萍
Owner NORTHWEST A & F UNIV
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