Method for simultaneously extracting microbial intracellular and extracellular DNAs (deoxyribonucleic acids) in sewage biological treatment water sample
A sewage biological treatment and microbial technology, applied in DNA preparation, recombinant DNA technology, etc., can solve the problems of extracting microbial intracellular and extracellular DNA at the same time, avoiding loss, good lysis effect and high yield. Effect
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Embodiment 1
[0033] When using a membrane bioreactor (membrane bioreactor, MBR) to treat domestic sewage, analyze the intracellular and extracellular DNA content of the water sample inside the membrane bioreactor.
[0034] Using 8L submerged hollow fiber membrane MBR, continuous aeration of 200L / h, HRT of 24h, sludge load of 0.24kg COD / kg VSS·d, stable operation at room temperature. Take 10mL of MBR internal mixed liquid for DNA extraction operation, the mixed liquid sample contains more activated sludge, and the color is light brown yellow.
[0035] Add 10mL NaH to the MBR mixture 2 PO 4 The solution (0.12M, pH 8.0) and 1g of polyvinylpyrrolidone (Polyvinylpolypyrrolidone, PVPP) were placed on a shaker (250rpm) at room temperature for 10 minutes to shake and mix well.
[0036] Then the sample was centrifuged (10000g, 4°C, 10min), and the centrifuged supernatant was further filtered with a sterile filter membrane (pore size 0.22μm), centrifuged and filtered to obtain the precipitate with...
Embodiment 2
[0040] When using a membrane bioreactor (membrane bioreactor, MBR) to treat domestic sewage, analyze the intracellular and extracellular DNA content of the membrane bioreactor effluent sample.
[0041] The difference between this example and Example 1 is that the sewage sample comes from the effluent of the reactor, the color is clearer, and the microbial content is less, so the sediment is obtained by centrifugation and filtration with NaH 2 PO 4 (0.12M, pH 8.0) after resuspension, repeat the operation of centrifugation and filtration only once, and use the freeze-thaw method to disrupt the cells, that is, alternately place the mixed solution in liquid nitrogen (1min) and 60°C water bath (20min) only once, other The operation was the same as in Example 1, and finally the content of intracellular DNA in the water sample was 0.091±0.013 μg / mL, the content of extracellular DNA was 0.064±0.032 μg / mL, and the ratios of absorbance at 260 nm to absorbance at 280 nm were 1.45 and 1.5...
Embodiment 3
[0043] Using the pure water sample as a blank control, the same steps as in Example 1 were used to extract the contents of intracellular and extracellular DNA to be 0.
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