Method for quantifying ammonia oxidizing archaea in lacustrine deposit

A technology for ammonia oxidizing archaea and sediments, applied in biochemical equipment and methods, microbial measurement/inspection, fluorescence/phosphorescence, etc., can solve the problem of long cycle, large error, inability to accurately reflect the number of ammonia oxidizing archaea, etc. problems to achieve the effect of accurate research methods

Inactive Publication Date: 2011-11-02
HOHAI UNIV
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Problems solved by technology

These traditional methods have a long period of time and large errors, and cannot accurately reflect the number of ammonia-oxidizing archaea in lake sediments

Method used

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  • Method for quantifying ammonia oxidizing archaea in lacustrine deposit
  • Method for quantifying ammonia oxidizing archaea in lacustrine deposit
  • Method for quantifying ammonia oxidizing archaea in lacustrine deposit

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[0020] For making the purpose of the present invention, technical scheme and advantage clearer, the present invention is further described as an example with the quantitative research of ammonia-oxidizing archaea in Taihu Lake sediment below:

[0021] This method for the quantitative study of ammonia-oxidizing archaea in sediments involves the following steps:

[0022] (1) Extraction of DNA

[0023] Lake sediment samples were collected and freeze-dried. Weigh 5 g of the lyophilized sediment sample into a 50 mL centrifuge tube, add 9 mL of sterile water, 2 mL of Tris-HCl buffer, 2.5 mL of EDTA solution and 5 mL of 10% SDS solution. After mixing, place in a water bath at 65°C for 60 min. Centrifuge at 12000 rpm for 10 min at room temperature. The supernatant was collected in a 50 mL centrifuge tube, 3 mL of 5M NaCl, 2 mL of 10% CTAB solution and 10 mL of proteinase K were added, mixed slightly, and incubated in a water bath at 65°C for 20 min. Add 16 mL of chloroform:isoamyl...

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Abstract

A method for quantifying ammonia oxidizing archaea in lacustrine deposit. The method comprises the following steps of (1) extracting DNA, and (2) constructing an ammonia oxidizing archaea plasmid standard substance and analyzing a sample, wherein the copy number of Ammonia oxidizing archaea in an unknown sample is obtained through processes of a detection of a Ct value of the unknown sample and a contrast of an amplification standard curve and the Ct value after the amplification standard curve is obtained. In the invention, ammonia oxidizing archaea in lacustrine deposit is detected quantificationally through a real-time fluorescent quantitative polymerase chain reaction (PCR) technology. The method is a rapid, simple and accurate method for researching a nitrogen cycle mechanism of lacustrine deposit.

Description

[0001] technical field [0002] The invention belongs to the technical field of environmental microbial ecology, and in particular relates to a method for quantitatively measuring ammonia-oxidizing archaea in lake sediments. [0003] Background technique [0004] At present, the eutrophication of freshwater lakes has become one of the important environmental problems facing our country. The excessive input of nutrients such as nitrogen and phosphorus is an important reason for the eutrophication of lakes. Therefore, studying the biogeochemical cycle of nitrogen in lake ecosystems can provide a basis for elucidating the formation mechanism of lake eutrophication and finding corresponding control measures. Lake sediments are an important part of the freshwater ecosystem, and are also the habitat of a large number of microbial groups. These microbial groups play an important role in the cycle of nutrients such as nitrogen and phosphorus in the lake, and play an important role ...

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Application Information

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IPC IPC(8): C12Q1/68C12Q1/06G01N21/64
Inventor 赵大勇马婷燕文明姜翠玲徐于楠黄睿王猛
Owner HOHAI UNIV
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