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Method and kit for rapidly extracting DNA of soil

A kit and soil technology, applied in the field of microbiology and molecular biology, can solve the problems of difficult removal of humus and low DNA recovery rate, and achieve the effects of reducing physical health damage, simple operation, and strong applicability

Inactive Publication Date: 2021-04-13
苏州易迈吉生物医药科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In order to solve the problems of difficult removal of humus in different soil samples and low DNA recovery rate in the process of extracting and purifying soil microbial DNA, the present invention provides a humus-free, high-purity, high-concentration extract from various soil samples within 20 minutes. Methods and kits for microbial genomic DNA

Method used

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  • Method and kit for rapidly extracting DNA of soil
  • Method and kit for rapidly extracting DNA of soil
  • Method and kit for rapidly extracting DNA of soil

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] (1) Weigh 0.5g of paddy field moist soil sample into a 2mL EP tube, add 200μL of Pre-Wash solution, and shake on a vortex shaker for 2min;

[0031] (2) Add 600 μL of soil lysate, vortex and shake for 5 minutes until the sample is fully mixed; incubate at 70°C for 10 minutes, shake 2-3 times during the incubation;

[0032] (3) Centrifuge at 12,000 rpm for 3 min. Take the supernatant to a new 1.5ml centrifuge tube;

[0033] (4) Transfer the supernatant obtained in step (3) to a new EP tube, then add 100 µL of impurity precipitation agent to the supernatant, vortex to mix, and then ice-bath for 5 min. Centrifuge at 12,000rpm for 3min;

[0034] (5) Transfer 500 µL of the supernatant obtained in step (4) to a new EP tube, add 250 µL of DNA binding solution and 500 µL of ethanol;

[0035] (6) Transfer the supernatant mixture obtained in step (5) to a DNA adsorption silica gel column, centrifuge at a speed of 12000rpm for 1min, remove the centrifugate and retain the DNA ads...

Embodiment 2

[0041] (1) Weigh 0.5 g of dry soil samples near chemical plants into a 2 mL EP tube, add 300 μL of Pre-Wash solution, and shake on a vortex shaker for 2 min;

[0042] (2) Add 700 μL of soil lysate, vortex and shake for 5 minutes until the sample is fully mixed; incubate at 70°C for 10 minutes, shake 2-3 times during the incubation;

[0043] (3) Centrifuge at 12,000 rpm for 3 min. Take the supernatant to a new 1.5ml centrifuge tube;

[0044](4) Transfer the supernatant obtained in step (3) to a new EP tube, then add 100 µL of impurity precipitation agent to the supernatant, vortex to mix, and then ice-bath for 5 min. Centrifuge at 12,000rpm for 3min;

[0045] (5) Transfer 500 µL of the supernatant obtained in step (4) to a new EP tube, add 250 µL DNA binding solution, 500 µL ethanol and 10 µL DNA adsorption magnetic beads, and vortex to mix;

[0046] (6) Place the EP tube in (5) on the magnetic stand for 1 min. After the magnetic beads are completely adsorbed, use a pipette ...

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Abstract

The invention relates to a method for efficiently extracting microorganism DNA from soil. A kit does not need an organic solvent, can effectively remove interference of humus, lipid and carbohydrates on DNA extraction by using a method of combining guanidine salt and an inorganic salt buffer solution DNA extraction kit, can adapt to various soil types, can effectively enrich DNA, and can remove pollution of humus. DNA purity and concentration detection experiment results prove that the DNA obtained by the extraction method can meet the requirements of downstream experiments, and reduce the influence of humus effectively.

Description

technical field [0001] The invention belongs to the technical field of microbiology and molecular biology, and in particular relates to a method and a kit for efficiently extracting soil microbial DNA. Background technique [0002] Due to the diversity of soil microenvironment, soil microorganisms are highly diverse. The study of these highly diverse microorganisms and community functions can not only understand how life tolerates extreme environments, new bioenergy, life evolution, and the interaction between microorganisms and the environment, but also can be used for new diagnosis of human diseases, new treatments, etc. method development. [0003] An effective means to study the diversity of soil microorganisms is to separate, purify and analyze the DNA in the soil, and track the behavior of certain target strains or recombinant genes in the natural environment; it can also be used to reveal the diversity of genes in the soil microbial ecosystem It is also a very criti...

Claims

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Application Information

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IPC IPC(8): C12N15/10
CPCC12N15/1006
Inventor 贾萌萌潘韵芝朱国强
Owner 苏州易迈吉生物医药科技有限公司
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