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A kind of method for extracting total dna of crape myrtle

A technology of crape myrtle and extracting solution, which is applied in the field of extracting crape myrtle total DNA, can solve problems such as the difficulty of extracting crape myrtle total DNA, and achieve the effects of avoiding tight combination, simple extraction, and solving difficult extraction

Inactive Publication Date: 2016-03-30
NORTHWEST A & F UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The purpose of the present invention is to provide a method for extracting total DNA of Lagerstroemia indica, which solves the problem of difficult extraction of Lagerstroemia indica total DNA

Method used

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  • A kind of method for extracting total dna of crape myrtle
  • A kind of method for extracting total dna of crape myrtle
  • A kind of method for extracting total dna of crape myrtle

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Lagerstroemia indica leaves were collected in Yangling, Shaanxi at the end of October 2103;

[0045] step 1:

[0046] Take the crape myrtle stem tissue out of the ultra-low temperature refrigerator at -80°C, weigh 1.5g, cut the tissue with scissors and keep the plant tissue in a frozen state to obtain the chopped plant tissue;

[0047] Step 2:

[0048] Add 0.3 g of PVP powder with a relative molecular mass of 45,000-55,000 directly to the shredded plant tissue in step 1, grind and crush the Lagerstroemia myrtle plant tissue in liquid nitrogen, and transfer the ground powder to five 1.5mL sterilized centrifuge tubes In each centrifuge tube, add 1 mL of CTAB extract preheated to 65 °C (CTAB extract consists of: 100 mM Tris-HCl at pH 8.0, 1.4 mol / L NaCl, 4% CTAB, 20 mM ethylenediamine at pH 8.0 Tetraacetic acid (EDTA) and 2% 2-mercaptoethanol, 2-mercaptoethanol should be added before use), kept in a water bath at 65°C for 60 minutes, and the centrifuge tube was shaken ev...

Embodiment 2

[0056] Lagerstroemia indica leaves were collected in Yangling, Shaanxi at the end of October 2103;

[0057] step 1:

[0058] Take out the Lagerstroemia indica leaf tissue from the ultra-low temperature refrigerator at -80°C, weigh 0.6g, cut the tissue with scissors and keep the plant tissue in a frozen state to obtain the chopped plant tissue;

[0059] Step 2:

[0060] Add 0.06g of PVP powder with a relative molecular mass of 45000-55000 directly to the shredded plant tissue in step 1, grind and crush the Lagerstroemia myrtle plant tissue in liquid nitrogen, and transfer the ground powder to five 1.5mL sterilized centrifuge tubes In each centrifuge tube, add 1 mL of CTAB extract preheated to 65 °C (CTAB extract consists of: 100 mM Tris-HCl at pH 8.0, 1.4 mol / L NaCl, 4% CTAB, 20 mM ethylenediamine at pH 8.0 Tetraacetic acid (EDTA) and 2% 2-mercaptoethanol, 2-mercaptoethanol was added before use), kept at 65°C for 80 minutes in a water bath, and the centrifuge tube was shaken ...

Embodiment 3

[0068] Lagerstroemia indica leaves were collected in Yangling, Shaanxi at the end of October 2103;

[0069] step 1:

[0070] Take the crape myrtle stem tissue out of the ultra-low temperature refrigerator at -80°C, weigh 1.5g, cut the tissue with scissors and keep the plant tissue in a frozen state to obtain the chopped plant tissue;

[0071] Step 2:

[0072] Add 0.6 g of PVP powder with a relative molecular mass of 45,000-55,000 directly to the shredded plant tissue in step 1, grind and crush the Lagerstroemia myrtle plant tissue in liquid nitrogen, and transfer the ground powder to five 1.5mL sterilized centrifuge tubes In each centrifuge tube, add 1 mL of CTAB extract preheated to 65°C (CTAB extract consists of: 100 mM Tris-HCl at pH 8.0, 1.4 mol / L NaCl, 4% CTAB, 20 mM EDTA at pH 8.0 and 2% 2-mercaptoethanol, add 2-mercaptoethanol before use), keep warm in a water bath at 65°C for 120 minutes, shake the centrifuge tube every 15 minutes during the water bath keep warm, so ...

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Abstract

The invention discloses a method of extracting total DNA of lagerstroemia indica. The method includes cutting lagerstroemia indica tissue into pieces at low temperature, adding polyvinylpyrrolidone (PVP) powder, grinding in liquid nitrogen, adding a cetyltrimethylammonium bromide (CTAB) extracting liquid, maintaining the temperature in a water bath, centrifuging at room temperature, adding organic solvent having the same volume as liquid supernatant into the liquid supernatant, centrifuging the mixture of the liquid supernatant and the solvent, depositing the liquid supernatant, centrifuging, cleaning, drying, and the like. According to the method, the PVP powder is added into the plant tissue in the extraction process, and the PVP powder can combine polysaccharides and polyphenols, thus avoiding joint connection of the polysaccharides with the DNA and oxidation of the polyphenols, and overcoming a problem of difficult extraction of the total DNA of the lagerstroemia indica. Extraction is simple, rapid and efficient. The method provides foundations for molecular biology study of the lagerstroemia indica and other plants rich in polysaccharides and polyphenols.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, and in particular relates to a method for extracting total DNA of crape myrtle. Background technique [0002] Lagerstroemia indica belongs to the genus Lagerstroemia in the family Lythraceae. Lagerstroemia indica is rich in polysaccharides, polyphenols and secondary metabolites, and extracting its DNA has always been a difficult problem. There are many methods for extracting polysaccharide and polyphenol plant DNA, but there is no report on the method successfully used to extract the genomic DNA of Lagerstroemia indica. At present, experts and scholars studying Lagerstroemia at home and abroad choose more finished kits when extracting Lagerstroemia indica DNA (Cai, M., Pan, H.T., Wang, X.F., He, D., Wang, X.Y., Wang, X.J., Zhang, Q.X., 2011 .DevelopmentofnovelmicrosatellitesinLagerstroemiaindicaandDNAfingerprintinginChineseLagerstroemiacultivars.Sci.Hortic.131,88–94.;Graham,S.A.,Hall,...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/10
Inventor 张荣孙广宇杜银银
Owner NORTHWEST A & F UNIV
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