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A high-efficiency extraction method of guava leaf genome dna

An extraction method and guava technology, applied in the field of molecular biology, can solve the problems of difficult to mention high-quality genomic DNA, can not be applied well, complex species, etc., improve the extraction rate and purity, and is conducive to popularization and utilization , a wide range of effects

Active Publication Date: 2020-10-16
THE SECOND XIANGYA HOSPITAL OF CENT SOUTH UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Traditional DNA extraction methods are not easy to obtain high-quality genomic DNA, and it takes a long time and the operation steps are cumbersome
The types of secondary metabolites in guava leaves are extremely complex, and the degree of heterogeneity of secondary metabolites in leaves of different leaf ages is high between different species and the same species in different seasons; therefore, the genome of a plant leaf DNA extraction methods are often not well applied to the extraction of genomic DNA from leaves of another plant, and it is particularly important to explore a fast, safe, cost-effective method for the extraction of genomic DNA from guava leaves

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] A high-efficiency extraction method of guava leaf genome DNA, comprising the following steps:

[0031] (1) Put 0.5 g of chopped guava leaves into a sterile mortar, add liquid nitrogen, use a high-frequency vibrating ultrafine powder machine to grind the guava leaves into powder, then transfer the powder into a centrifuge tube, add 1.2mL of washing liquid, after mixing, ultrasonic treatment, let stand in ice for 15min, centrifuge at 10000rpm for 10min to obtain supernatant a and bottom residue a, wherein, the composition of washing liquid includes: 50mM Tris-HCl, 5mM EDTA, 350mM sorbitol, The volume fraction is 2% PVP, the volume fraction is 0.5% β-mercaptoethanol, and the pH value of the control lotion is 8.0;

[0032] (2) Add 50 μL of cellulase and pectinase to the bottom residue a, and after standing in ice for 10 minutes, centrifuge at 10,000 rpm / min for 10 minutes to obtain supernatant b and bottom residue b;

[0033](3) Add CTAB extraction buffer to the bottom res...

Embodiment 2

[0040] A high-efficiency extraction method of guava leaf genome DNA, comprising the following steps:

[0041] (1) Put 0.5 g of chopped guava leaves into a sterile mortar, add liquid nitrogen, use a high-frequency vibrating ultrafine powder machine to grind the guava leaves into powder, then transfer the powder into a centrifuge tube, add 1.0mL of washing liquid, after mixing, ultrasonic treatment, let stand in ice for 15min, centrifuge at 10000rpm for 10min to obtain supernatant a and bottom residue a, wherein, the composition of washing liquid includes: 50mM Tris-HCl, 5mM EDTA, 350mM sorbitol, The volume fraction is 2% PVP, the volume fraction is 0.5% β-mercaptoethanol, and the pH value of the control lotion is 8.0;

[0042] (2) Add 55 μL of cellulase and pectinase to the bottom residue a, and after standing in ice for 10 minutes, centrifuge at 10,000 rpm / min for 10 minutes to obtain supernatant b and bottom residue b;

[0043] (3) Add CTAB extraction buffer to the bottom re...

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PUM

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Abstract

The invention discloses an efficient extraction method of guava leaf genome DNA (deoxyribonucleic acid). The method includes the steps: (1) adding liquid nitrogen into guava leaves, grinding the guavaleaves into powder, adding washing liquor again, ultrasonically treating mixture, standing the mixture in an ice, and centrifuging the mixture to obtain liquid supernatant a and bottom slag a; (2) adding an enzyme into the bottom slag a, standing mixture in an ice, and centrifuging the mixture to obtain liquid supernatant b and bottom slag b; (3) adding CTAB (cetyl trimethyl ammonium bromide) extraction buffer liquid into the bottom slag b, and centrifuging mixture to obtain liquid supernatant c and bottom slag c; (4) adding Tris saturated phenol, chloroform and isoamyl alcohol into the liquid supernatant c, and centrifuging mixture to obtain liquid supernatant d and bottom slag d; (5) adding isopropyl alcohol into the liquid supernatant d, and centrifuging mixture to obtain liquid supernatant e and bottom slag e; (6) washing the bottom slag e acquired in the step (4) by ethyl alcohol, centrifuging mixture to collect bottom slag to obtain the guava leaf genome DNA. The guava leaf genome DNA is high in purity and extraction rate.

Description

technical field [0001] The invention belongs to the technical field of molecular biology, in particular to a method for efficiently extracting guava genome DNA. Background technique [0002] Guava is a plant of the Myrtaceae Guava genus. The shape of guava fruit is spherical, oval, oval and pear-shaped. The peel is generally green, red, and yellow, and the flesh is white, red, and yellow. The meat is very soft, the gravy is rich, and the taste is sweet. 8%-11%, rich in potassium, iron, carotene and other substances, extremely rich in nutrition, it is the best fruit for beauty and weight loss. Its fruit can not only be eaten fresh, but also can be processed into fruit juice, jam, preserved fruit, and can also be made into bonsai. It has broad market prospects and is currently one of the best-selling fruits in Hong Kong, Macao, Taiwan and Southeast Asia. In addition, guava leaves and young fruit slices are dried and made into tea, which can assist in the treatment of diabete...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/10
CPCC12N15/1003
Inventor 谢志国申丽周智广王俊俊曾伟民胡芳李交昆余润兰吴学玲刘元东吴晨晨李芳刘阿娟邱冠周
Owner THE SECOND XIANGYA HOSPITAL OF CENT SOUTH UNIV
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