Method for extracting DNA from dry apricot leaf

A leaf and drying technology, applied in the field of DNA extraction, can solve the problems of difficulty in obtaining high-quality apricot leaves, affecting PCR amplification, genetic polymorphism analysis research, and great differences in species and content, etc., to improve the extraction purity and yield The effect of high yield, good quality of extracted DNA, and high yield

Inactive Publication Date: 2015-04-22
TARIM UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The types and contents of secondary metabolites in plants vary greatly, and sometimes the types and contents of secondary metabolites in different states of the same plant organ or tissue are also different. The DNA extraction method optimized for a certain species may not be applicable to other species, so it is difficult to obtain DNA from high-quality apricot leaves
The traditi

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  • Method for extracting DNA from dry apricot leaf
  • Method for extracting DNA from dry apricot leaf
  • Method for extracting DNA from dry apricot leaf

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Embodiment 1

[0019] A method for extracting DNA from dry apricot leaves, comprising the following steps:

[0020] (1) Remove the petiole of the apricot leaf, weigh 20 mg of the leaf, add 0.001 g of PVP-40T, add liquid nitrogen, grind the leaf into a fine powder, and add the fine powder to a 2.0 mL centrifuge tube;

[0021] (2) Add 1mL of pre-cooled buffer extract solution, mix well and then ice bath for 15 minutes, invert and mix 2-3 times during the ice bath;

[0022] (3) Centrifuge at 7000g for 10min, discard the supernatant;

[0023] (4) Repeat steps (2) and (3) until the supernatant is not viscous;

[0024] (5) Quickly add 0.8mL of CTAB Buffer preheated to boiling to each tube, mix well, bathe in 65℃ water for 0.5-1h, and shake once every 15min;

[0025] (6) Add 0.01mL 100mg / L RNase, bathe in 37℃ water for 30-60min;

[0026] (7) Take it out and cool to room temperature, add 0.8mL of phenol:chloroform:isoamyl alcohol with a volume ratio of 25:24:1 to each tube, store at 4°C, gently i...

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Abstract

The invention belongs to the technical field of preparation of plant DNA and relates to a method for extracting DNA from a dry apricot leaf. The method comprises the following steps: firstly extracting secondary metabolite substances from the leaf; putting the grinded dry apricot leaf in an ice bath by using a precooling buffer extract solution, centrifuging, removing the supernatant liquid, and repeatedly extracting for a plurality of times to remove the secondary metabolite substances such as polysaccharide, polyphenol and ester from the apricot leaf before the DNA is released so as to prevent the secondary metabolite substances from forming a compound with the DNA; during liquid nitrogen grinding and treatment of the buffer extraction solution, adding a proper amount of PVP-40T to protect the DNA from oxidation and promote cell disruption so that more DNA can be released; adding a proper amount of RNA enzyme after the cell disruption so that RNA is removed and the introduced protease is removed as well; and extracting by using a proper amount of a mixed solution of phenol, chloroform and isoamylol by adopting an improved CTAB method to remove a great amount of protein so that the purity and quality of the DNA in the apricot leaf are improved. Compared with a generally-used method, the method disclosed by the invention is low in cost.

Description

technical field [0001] The invention belongs to the technical field of plant total DNA preparation, in particular to a method for extracting DNA from dried apricot leaves. technical background [0002] Plant leaves collected in the wild have to be transported and preserved over long distances. Regardless of young leaves or mature leaves, after drying through silica gel, although the leaves lose water rapidly, the plant leaf tissues still contain a large amount of polysaccharides, polyphenols, and proteins. and esters and other plant secondary metabolites and degraded DNA, which makes the yield, quality and difficulty of extracting DNA very different from those extracted from fresh leaf materials. [0003] DNA is the carrier of genetic information and the basic genetic material of plants. High-quality DNA samples are the material basis for molecular biology research such as restriction enzyme digestion, PCR amplification, molecular hybridization, genetic polymorphism analysi...

Claims

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Application Information

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IPC IPC(8): C12N15/10
Inventor 郭玲周慧杰罗华平
Owner TARIM UNIV
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