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Method for extracting nucleic acid from activated carbon biological membranes in drinking water treatment

A technology of drinking water treatment and extraction method, which is applied in the field of advanced treatment of drinking water, can solve problems such as difficult elution, and achieve the effect of simple operation

Inactive Publication Date: 2012-10-10
TSINGHUA UNIV
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Problems solved by technology

[0006] The purpose of the present invention is to solve the problem of difficult elution in the prior art due to the low biomass on the biological activated carbon and the tight combination with the carrier particles, reduce the adverse effect of humic acid on the extraction of microbial nucleic acid, and through the decompression treatment and ultrasonic elution method, Provides a method for effectively extracting microbial nucleic acids on bioactivated carbon to obtain DNA fragments suitable for downstream molecular biology operations in yield and purity

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  • Method for extracting nucleic acid from activated carbon biological membranes in drinking water treatment
  • Method for extracting nucleic acid from activated carbon biological membranes in drinking water treatment
  • Method for extracting nucleic acid from activated carbon biological membranes in drinking water treatment

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Embodiment

[0024] The biological activated carbon sample was taken from a waterworks in Beijing. In Examples 1-3, NaOH solution (50-200mmol / L) was used as the decomposing buffer, and the ultrasonic elution time was 1min, 2min and 5min, respectively, to extract DNA; Example 4 ~6 Use NaOH (50~200mmol / L)+PVP (PVP:NaOH 5:2, w / w) mixed solution as the decomposing buffer, and the ultrasonic elution time is 1min, 2min and 5min, respectively, to extract DNA.

[0025] The DNA extraction results are shown in Table 1, and the obtained total DNA A 260 / A 280 Values ​​above 1.6 indicate that there is no protein contamination in the total DNA, and some samples above 1.8 indicate partial RNA contamination, which can be removed by adding a little RNase. Using NaOH solution (100mmol / L) as the decomposing buffer, the amount of DNA extracted increased when the ultrasonic time was increased from 1 min to 5 min. Using NaOH (200mmol / L)+PVP (PVP:NaOH 5:2, w / w) mixed solution as the decomposing buffer, the DN...

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Abstract

The invention discloses a method for extracting nucleic acid from activated carbon biological membranes in drinking water treatment and relates to the technical field of microbiology of biological activated carbon process units in environmental engineering drinking water deep treatment. The method includes: firstly, subjecting activated carbon particles to dehumification by the aid of dehumification buffer solution; and then performing water bath and ultrasonic elution. The method specifically includes: subjecting activated carbon to dehumification by the aid of the dehumification buffer solution obtained by mixing NaOH solution or NaOH with polyvinylpyrrolidone, and extracting the microbial nucleic acid on the biological activated carbon after ultrasonic elution and chemical cleavage. The total DNA (deoxyribonucleic acid) length is larger than 20 Kb so as to completely meet the requirements of PCR (polymerase chain reaction) analysis. According to the method, the problem of difficulty in elution resulted from low biomass of the biological activated carbon and close combination of the biological activated carbon with carrier particles is solved, and adverse effect of humic acid on microbial nucleic acid extraction is weakened, so that microbial cells are sufficient in cleavage. The method is simple and convenient to operate and high in DNA output and purity.

Description

technical field [0001] The invention relates to a method for extracting microbial nucleic acid from biological activated carbon in drinking water treatment, and belongs to the technical field of advanced treatment of drinking water. Background technique [0002] Activated carbon has good adsorption performance and stable chemical properties, can withstand strong acid and strong alkali, can withstand water immersion and high temperature, is lighter than water, and is a porous hydrophobic adsorbent, so it has been widely used to remove odors in drinking water , chroma, natural and synthetic organic matter. The biofilm attached to activated carbon, mainly composed of microorganisms and their extracellular polymers, is the main part of the biodegradation of harmful substances in water. Therefore, the identification and analysis of the microbial community structure and distribution on the surface of activated carbon is of great significance for clarifying the biodegradation mech...

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
Inventor 刘文君张明露王占朝余祎
Owner TSINGHUA UNIV
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