A method for extracting total DNA from environmental samples with high efficiency in removing humus
An environmental sample and extraction method technology, which is applied in the field of total DNA extraction of environmental samples for efficiently removing humus, can solve the problems of needing purification steps, low purity of soil total DNA, time-consuming and labor-intensive, etc., and achieves the effect of improving the extraction purity.
Inactive Publication Date: 2011-12-21
INNER MONGOLIA UNIVERSITY
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The above method directly extracts the total DNA from the soil with low purity and requires subsequent purification steps, which is time-consuming and labor-intensive.
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[0019] The present invention will be described in detail below in conjunction with specific embodiments.
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The invention discloses a method for extracting total DNA from an environmental sample with high efficiency for removing humus, comprising the following steps: (1) taking 0.3 g of sieved soil sample in a 2 mL centrifuge tube equipped with three 2.5 mm-3.0 mm sterilized glass beads, Add 1 mL of decomposing buffer, Vortex-Genie2 (Mobio Laboratories Inc) for 1 min, centrifuge at 12000 r/min for 2 min, discard the supernatant; (2) add 1 mL of 0.5 mol/L calcium chloride solution, Vortex-Genie2 (Mobio Laboratories Inc) ) Vortex for 2 min, centrifuge at 12,000 r/min for 1 min, and discard the supernatant; (3) Add 800 μL of DNA extraction buffer, and Vortex-Genie2 (Mobio Laboratories Inc) vortex for 5 s. Rapid and efficient extraction of total soil DNA, providing high-purity genomic DNA for soil metagenomics research.
Description
technical field [0001] The invention relates to a method for extracting total DNA of an environmental sample for efficiently removing humus, and belongs to the field of biotechnology. Background technique [0002] Soil genomic DNA extraction is the basis of environmental metagenomics research. The extraction of soil DNA can be achieved in two ways. One is to separate the microbial cells in the soil from soil particles before lysing the cells, and then lyse the cells to obtain the total DNA; the other is to directly lyse the cells and extract the total DNA from the soil. , the total DNA yield obtained by this approach is large, but the purity is low, mainly due to the impact of impurities such as humic acid. [0003] At present, there are many methods for soil DNA extraction, such as liquid nitrogen grinding method, microwave crushing method, glass bead cracking method, enzymatic cracking method, rapid freeze-thaw method, ion exchange method, solvent-glass bead mixing method...
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IPC IPC(8): C12N15/10
Inventor 赵吉李靖宇于景丽武琳慧
Owner INNER MONGOLIA UNIVERSITY
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