Extraction method of plant genome DNA rich in polysaccharides

A plant genome and extraction method technology, which is applied in the field of plant genome DNA extraction, can solve the problems of expensive kits, waste of experiment costs, and low DNA purity, and achieve good extraction effects and good gene integrity

Pending Publication Date: 2019-03-08
SHANGHAI PASSION BIOTECHNOLOGY CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

This technology has particularly high requirements on the quality and purity of DNA. For example, DNA degradation will result in less data, poor data quality, and unsatisfactory splicing effect; DNA purity is not high, and contamination by sugar or salt will cause polymerase activity to decr

Method used

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  • Extraction method of plant genome DNA rich in polysaccharides
  • Extraction method of plant genome DNA rich in polysaccharides
  • Extraction method of plant genome DNA rich in polysaccharides

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[0036] The present invention will be further described by the following examples, but these examples should not be used to explain the limitation of the present invention.

[0037] Samples of jasmine petals, oak leaves, crape myrtle leaves, catalpa leaves, and cloves are processed according to the following steps and then put on the machine:

[0038] 1. Add 1ml pre-cooled CTAB-free buffer, 20ul mercaptoethanol, shake and mix well;

[0039] 2. Place on ice for 2-3 minutes, centrifuge at 4°C, 4000 rpm for 10 minutes, and discard the supernatant;

[0040] 3. Add 1ml PBS, mix well, centrifuge at 4°C, 4000rpm for 10min, discard the supernatant;

[0041] 4. Choose to wash with PBS again according to the viscosity, so as to achieve the removal of polysaccharide impurities;

[0042] 5. Add 700ul of SDS solution preheated at 65°C, 10ul of mercaptoethanol, and mix well;

[0043] 6. Put it in a 65°C water bath for 25 minutes, and mix it by inverting at least 3 times during the period;...

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Abstract

The invention discloses an extraction method of plant genome DNA rich in polysaccharides. The method comprises the following steps: grinding through liquid nitrogen; adding precooled CATB-free bufferand mercaptoethanol, and sufficiently and uniformly vibrating; laying the materials on ice, controlling the temperature to be 4 DEC C, performing centrifugation, and abandoning supernatant; washing according through PBS according to viscosity, adding an SDS solution and mercaptoethanol, and uniformly mixing; then performing water bath, and performing reversing and mixing during water bath; addingequivoluminal phenol and chloroform after the materials are cooled, and reversing and mixing; performing centrifugation, taking supernatant, adding RNase A, performing reversing and mixing, and layingthe materials at a room temperature; adding isopropanol, reversing and mixing, laying the materials at a room temperature or a low temperature, performing centrifugation, abandoning supernatant, washing through 75 percent ethanol, and adding an appropriate volume of sterile water for dissolving. The method has a better extraction effect on plant gene rich in polysaccharides, and compared with conventional methods, the method has the advantages that the gene integrity and subsequent purity are better.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for extracting polysaccharide-rich plant genome DNA. Background technique [0002] In recent years, the development of molecular biology technology and sequencing technology has provided a good foundation for researchers to study microbial genome sequences and the synthesis mechanism of secondary metabolites, and the acquisition of plant DNA is the basis of these work. [0003] Polysaccharides exist in large quantities in plant cells as structural constituents and nutrient storage substances of plant cells. Polysaccharides are the main substances that interfere with plant DNA extraction. Many physical and chemical properties of polysaccharides are very similar to DNA, so it is difficult to separate them. The colloidal substance formed by the combination of polysaccharide and DNA not only affects the operation of the experiment, but also inhibits the activity of many enzyme...

Claims

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Application Information

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IPC IPC(8): C12N15/10
CPCC12N15/1003
Inventor 刘艳艳马义峰张洪波孙子奎
Owner SHANGHAI PASSION BIOTECHNOLOGY CO LTD
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