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Extraction method of DNAs from human body skin bacterium groups

A human skin and extraction method technology, applied in the field of DNA extraction, can solve the problems of high price, high sample quality requirements, cumbersome extraction steps, etc., and achieve the effect of simple operation, superior effect and low price

Inactive Publication Date: 2020-05-08
QILU CHILDRENS HOSPITAL OF SHANDONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Existing skin flora DNA is mostly extracted with commercial kits, but the special kits for skin flora are expensive, the extraction steps are cumbersome, and the sample quality requirements are high. Therefore, a simple, low-cost, and superior-effect human body Skin flora DNA extraction method is very necessary

Method used

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  • Extraction method of DNAs from human body skin bacterium groups
  • Extraction method of DNAs from human body skin bacterium groups
  • Extraction method of DNAs from human body skin bacterium groups

Examples

Experimental program
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Effect test

Embodiment

[0083] After taking the skin swab, store the cotton swab in the bacterial preservation solution, and vortex for 10 seconds at the maximum gear of the vortex oscillator to completely detach the bacteria from the swab;

[0084] Take 600ul of bacterial liquid in a sterile 1.5mL centrifuge tube, centrifuge at 12000rpm for 15min, make all the bacteria settle at the bottom of the tube, and remove as much liquid as possible;

[0085] Add 300ul of 2X CTAB solution preheated at 65°C to the tube, vortex at the maximum gear for 10S, then heat in a metal bath at 95°C for 10min;

[0086] Add 300ul phenol-form mixture solution (saturated phenol: chloroform: isoamyl alcohol = 25: 24: 1) to the tube and vortex at the maximum gear to fully mix the solution and complete protein extraction;

[0087] Centrifuge at 12000rpm for 5min, carefully transfer 200ul supernatant to a new sterile 1.5mL centrifuge tube, add 3 times the volume (600ul) of Buffer B2 (SanPrep Column DNA Gel Recovery Kit), and in...

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Abstract

The application provides an extraction method of DNAs from human body skin bacterium groups. According to the extraction method, a CTAB-phenol imitating method is organically combined with a reagent kit, a compound is formed through CTAB, protein and polysaccharid, phenol imitating mixed liquor is used for extraction and removal, DNAs are left behind, repeated washing is performed through an adsorption column of the reagent kit, impurities are removed, high-quality DNAs are obtained, and subsequent library construction and high-flux sequencing requirements are met. Compared with an existing reagent kit for extracting total DNA from skin bacterium groups, the extraction method disclosed by the invention is simple and convenient to operate, and special instrument and equipment and fittings are not needed; and the extracted DNAs can be subjected to PCR amplification to obtain accurate objective strips, and besides, concentration and purity of the DNAs can meet high-flux sequencing requirements.

Description

technical field [0001] The present application relates to the technical field of DNA extraction, in particular to a method for extracting human skin flora DNA. Background technique [0002] Many microorganisms invisible to the naked eye live on the surface of the human skin, and these microorganisms are closely related to human health and diseases, especially to the health of the skin. [0003] In order to study which types of microorganisms exist in the skin and affect human health, after swabbing the skin with a cotton swab, extracting the DNA of the skin flora is the most critical step, which is directly related to the subsequent sequencing and analysis results. Existing skin flora DNA is mostly extracted with commercial kits, but the special kits for skin flora are expensive, the extraction steps are cumbersome, and the sample quality requirements are high. Therefore, a simple, low-cost, and superior-effect human body Skin flora DNA extraction method is very necessary. ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6806
CPCC12Q1/6806C12Q2523/308
Inventor 盖中涛赵璐葛永胜刘毅
Owner QILU CHILDRENS HOSPITAL OF SHANDONG UNIV
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