Unlock instant, AI-driven research and patent intelligence for your innovation.

Method for extracting microbial genome DNA in complex sample

An extraction method and microbial technology, applied in the biological field, can solve problems such as limited space for improvement, and achieve the effects of cost saving, low cost and good environmental protection effect

Inactive Publication Date: 2022-02-25
GENFINE BIOTECH BEIJING CO LTD
View PDF5 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, the removal of inhibitory organic substances such as humic acids mainly relies on the combination of chemical reagents in the solution and organic substances such as humic acid to form precipitation under certain conditions to achieve the removal effect. After a certain level, the room for further improvement is very limited

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Method for extracting microbial genome DNA in complex sample
  • Method for extracting microbial genome DNA in complex sample
  • Method for extracting microbial genome DNA in complex sample

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Embodiment 1, the DNA extraction of soil sample

[0043] 1) Preliminary lysis of microbial cells: Take about 300mg of soil samples and mix them with 250mg of grinding beads, put them into a 2ml screw-cap centrifuge tube containing 200uL extraction buffer Ⅰ and 800 uL extraction buffer Ⅱ, then add 10 uL to a concentration of 10mg / ml RNase A; after adding, place the centrifuge tube on a tissue grinder and vibrate for 15 minutes.

[0044] 2) Further lyse microbial cells: add 10 mg lysozyme to the preliminary lysate and mix well, incubate at 50°C for 1 hour, then add 100 uL of 20% SDS solution, mix well, and incubate at 70°C for 30 minutes, centrifuge at 12000rpm for 10 minutes, and collect the supernatant.

[0045]3) Preparation of activated carbon filter column: Weigh 200 mg of activated carbon particles (150-200 mesh size), put it into the filter column used in the nucleic acid extraction process, take 500 uL of sterilized deionized water into the filter column, 8000rpm...

Embodiment 2

[0054] Embodiment 2, the DNA extraction of stool sample

[0055] 1) Preliminary lysis of microbial cells: Take about 200 uL of liquid feces samples (preserved in feces preservation solution), mix them with 250 mg of grinding beads, put them into a 2ml screw cap centrifuge containing 200 uL of extraction buffer Ⅰ and 800 uL of extraction buffer Ⅱ Into the tube, add 10 uL of RNase A with a concentration of 10mg / ml; after the addition, place the centrifuge tube on a tissue grinder and shake and grind for 5 minutes.

[0056] 2) Further lyse the microbial cells: incubate at 70°C for 30 minutes, centrifuge at 10,000 rpm for 10 minutes, and collect the supernatant.

[0057] 3) Preparation of activated carbon filter column: Weigh 200 mg of activated carbon particles (150-200 mesh size), put it into the filter column used in the nucleic acid extraction process, take 500 uL of sterilized deionized water into the filter column, 8000rpm Centrifuge for 5 minutes to prepare an activated ca...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
particle sizeaaaaaaaaaa
Login to View More

Abstract

The invention discloses a method for extracting microbial genome DNA in a complex sample. The method comprises the following steps: S1, cracking a sample to obtain supernate, wherein the sample is soil or excrement; S2, adding the supernate into an activated carbon filter column, performing centrifugal separation, and collecting filtrate; and S3, carrying out magnetic bead purification on the filtrate to realize extraction of the microbial genome DNA. According to the method disclosed by the invention, organic matter such as humic acid in the process of extracting the microbial genome DNA of the sample (soil and excrement) are removed through the activated carbon filter column, and purification can be realized in a centrifugal manner, so that the preparation cost of a humic acid removal solution and the low-temperature standing time during use are saved. The activated carbon adopted by the method disclosed by the invention is low in cost and is a renewable resource, so that the use of chemical reagents can be avoided, and a better environment-friendly effect is generated.

Description

technical field [0001] The invention relates to a method for extracting microbial genome DNA from complex samples, belonging to the field of biotechnology. Background technique [0002] Soil microorganisms are not only the most active and important part of the soil ecosystem, but also play a vital role in maintaining soil ecological functions. Soil microbes are very sensitive to changes in soil quality. Therefore, soil microbes have always been considered as one of the biological indicators most closely related to soil quality and sustainable development; scientific data on the composition and changes of their population structure are useful for studying changes in soil quality and predicting the impact of human activities on the soil environment. Damage and its potential risks provide important evidence. There are extremely rich microbial species in the natural ecological environment, but 85% to 99.9% of them are difficult to achieve recultivation under artificial control...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
CPCC12N15/1017C12N15/1013
Inventor 焦晓明韩典霖俞萍
Owner GENFINE BIOTECH BEIJING CO LTD