Extraction method of metagenome of microbes in seawater

A technology of metagenomics and extraction methods, which is applied in the field of extraction of seawater microbial metagenomics, can solve problems such as the difficulty of extracting DNA, achieve the effects of improving impurity interference, speeding up experiment speed, and increasing DNA concentration

Active Publication Date: 2016-10-26
厦门基源医疗科技有限公司
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And the types of seawater microorganisms in different waters are differe

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  • Extraction method of metagenome of microbes in seawater
  • Extraction method of metagenome of microbes in seawater

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[0015] Example

[0016] A method for extracting seawater microbial metagenomes. First, seawater samples are obtained and pretreated, and then the microbial metagenomes are extracted. The specific steps include:

[0017] (1) Removal of impurities in seawater: α-cellulose was added to the seawater sample at a mass-volume ratio of 2%, and allowed to stand at room temperature for 0.5 h; Use the negative pressure pump, pour the seawater sample above the filter, turn on the power supply of the negative pressure pump, and perform filtration; then replace the 0.2 um filter membrane to filter the filtrate again; collect both filter membranes; Quickly placed in liquid nitrogen for 1 min, and then placed at room temperature for 90 s, repeated freezing and thawing 5 times;

[0018] (2) Lysis of seawater microorganisms: cut the filter membrane and place it in a 5 ml centrifuge tube, then add 250 μl of cell lysate, 40 μl of lysozyme at a concentration of 20 mg / ml and 10 μl of EDTA at a co...

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Abstract

The invention provides an extraction method of metagenome of microbes in seawater. According to the invention, the special sample, seawater, undergoes saline ions removal treatment, double-membrane filtration and recovery processing. The invention aims to provide an efficient and rapid method for extracting metagenome of microbes in seawater. The method comprises the following specific steps: removal of impurities in seawater, cracking of microbes in seawater, dissociation and adsorption of DNA and DNA elution. Interference of many saline ions in seawater can be effectively eliminated; and microbial cells in seawater are fully cracked to fully release DNA. According to the DNA extracted by the method, A260/280 ratio of the DNA is maintained between 1.8 and 2.0. The obtained DNA has good quality and high purity, can meet requirements of gene library construction and qPCR and high-throughput sequencing, and provides good foundation for the research on heritable variation diversity of microbial genomes.

Description

technical field [0001] The invention provides a method for extracting seawater microbial metagenome, which belongs to the technical field of molecular biology. Background technique [0002] Seawater microorganisms are an important source of biologically active substances. The particularity of the marine environment and the complex interactions between marine organisms endow marine microorganisms with metabolic pathways and adaptation mechanisms that are different from terrestrial organisms, resulting in new genetic specificity and the synthesis of secondary organisms with unique structures and specific biological activities. metabolite. These active substances with abnormal structures and special effects are unmatched by terrestrial microorganisms. Since the 20th century, people have been using the method of isolation and culture to study the diversity of seawater microorganisms. That is to separate and purify seawater microorganisms from the environment, and then conduct...

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Application Information

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IPC IPC(8): C12N15/10
CPCC12N15/101
Inventor 李珊谢文龙刘青青王松林陈荣山肖辛野李奇渊姚迅
Owner 厦门基源医疗科技有限公司
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