Kit for extracting silt microbe genome DNA (deoxyribonucleic acid) on basis of magnetic bead process

A microorganism and genome technology, applied in the biological field, can solve the problem of not accounting for environmental microorganisms, and achieve the effect of shortening the extraction process and easy operation.

Inactive Publication Date: 2015-11-04
FUJIAN NORMAL UNIV
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Problems solved by technology

However, the bacteria that can grow on the medium only account for less than 1% of the total amount of environmental microorganisms

Method used

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  • Kit for extracting silt microbe genome DNA (deoxyribonucleic acid) on basis of magnetic bead process

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Embodiment Construction

[0021] figure 1 Among them, lanes 1 and 2 are the sludge microbial genomes extracted by this kit, and lane 3 is DL4500Marker from TAKARA.

[0022] A kit for extracting sludge microbial genomic DNA based on the magnetic bead method, the specific steps are as follows:

[0023] (1) Disintegration of sludge microbial cells: Weigh 1.5g of sludge sample into a 10ml centrifuge tube, add 4ml of Buffer A solution, shake for 5 minutes to mix, add lysozyme to a final concentration of 3mg / ml, shake for 5 minutes to mix. Shake at 37°C for 1 hour on a shaker at 225 r / min, add 600 μl of Buffer B solution, shake for 5 minutes and mix well, then bathe in water at 65°C for 2 hours, shake and mix for 1 minute every 10 minutes during this period,

[0024] (2) Extraction of impurity proteins: add 4ml Buffer C to the solution, shake the oscillator for 30-60s, centrifuge at 12000-13000rpm / min for 10-20min, transfer the supernatant to a new centrifuge tube, add an equal volume of Buffer D, centrifu...

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Abstract

The invention provides a kit for extracting silt microbe genome DNA (deoxyribonucleic acid) on the basis of a magnetic bead process. The method comprises the following steps: crushing silt microbe cells, removing impure proteins, recovering the microbe genome by a magnetic bead process to obtain the silt microbe genome DNA, and carrying out electrophoresis detection in 1% sepharose gel. The kit is mainly developed on the principle that the silyl magnetic beads specifically adsorb the genome DNA, is simple to operate, and greatly shortens the conventional silt microbe genome DNA extraction process, is more convenient and faster than the conventional silt microbe genome DNA extraction method, can implement large-scale extraction of the silt microbe genome DNA, and has certain economic value.

Description

technical field [0001] The invention relates to a kit for extracting genome DNA of sludge microorganisms based on a magnetic bead method, and belongs to the field of biotechnology. technical background [0002] Sludge is rich in microorganisms, and different microorganisms have different values. Studies have shown that in sludge, microbial communities work together to decompose organic compounds with larger molecular weight and more complex structure into the form of acetate. The Geobacters then transfer electrons from the acetate to electrodes that generate electricity. Some Geobacters are capable of converting toxic organics such as toluene into electrical charges, but a deeper understanding of how microbes generate electricity, and what other microbes do, will require studying them using genomics. [0003] The traditional method of isolating and cultivating microorganisms is the earliest method used in the study of sludge microbial diversity. However, the bacteria that ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
Inventor 王清水余彦
Owner FUJIAN NORMAL UNIV
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