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Collection and normal temperature drying storage method of water environment DNA samples

A storage method, drying technology at room temperature, applied in biochemical equipment and methods, determination/inspection of microorganisms, etc., can solve the problems of poor timeliness, high cost, complicated operation, etc., to prevent reproduction and growth and effectively kill active microorganisms. Effect

Pending Publication Date: 2020-09-08
南京易基诺环保科技有限公司
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  • Summary
  • Abstract
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  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Aiming at the shortcomings of cumbersome operation, high cost and poor timeliness in the preservation of collected environmental DNA samples by "freeze-drying" in the prior art, as well as the defects of monitoring interference caused by adding stabilizers, the present invention provides a water environment The method of DNA sample collection and normal temperature dry storage is to use a filter membrane to filter samples to enrich the environmental DNA on the filter membrane, then use isopropanol and ethanol to dry the filter membrane in turn, and finally dry the filter membrane for storage. The method can effectively prolong the storage time of environmental samples, reduce the storage conditions of environmental water samples, realize long-term storage of environmental DNA samples in the filter membrane under normal temperature conditions, significantly reduce costs, improve timeliness, and will not cause damage to subsequent DNA monitoring. interference

Method used

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  • Collection and normal temperature drying storage method of water environment DNA samples
  • Collection and normal temperature drying storage method of water environment DNA samples
  • Collection and normal temperature drying storage method of water environment DNA samples

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] In this embodiment, the water sample is filtered by a negative pressure filter device, the negative pressure filter device includes a negative pressure pump, and the negative pressure pump communicates with the water environment DNA sample filter cup with a filter membrane through a communication pipeline. A pressure relief valve is provided on the connecting pipeline.

[0043] Water environmental DNA sample testing Water samples were collected in Jiuxiang River (32.113681 north latitude, 118.945127 east longitude).

[0044] The filter membrane adopts Jinteng water system filter membrane, the pore size of the filter membrane is 0.45μm, and the diameter of the filter membrane is 50mm.

[0045] In this embodiment, the method steps for collecting and storing the water environment DNA samples are as follows:

[0046] (1) Use the negative pressure filter device to filter the water sample. After the water sample is filtered, turn off the negative pressure pump and open the pre...

Embodiment 2

[0051] In this embodiment, the water sample is filtered using a negative pressure filter device, which includes a negative pressure pump, and the negative pressure pump passes through a pipeline (which is provided with a pressure relief valve) and a water environment DNA with a filter membrane. The sample filter cup is connected.

[0052] The test water samples were collected in Jiuxiang River (32.113681 north latitude, 118.945127 east longitude).

[0053] The filter adopts Jinteng water system filter membrane, the filter membrane pore size is 0.45μm, and the filter membrane diameter is 50mm.

[0054] In this embodiment, the steps of the dry storage method for the DNA samples in the water environment are as follows:

[0055] (1) Use the negative pressure filter device to filter the water sample. After the water sample is filtered, turn off the negative pressure pump and open the pressure relief valve;

[0056] (2) After the pressure is zero, close the pressure relief valve, ...

Embodiment 3

[0060] In this embodiment, the water sample is filtered using a negative pressure filter device, which includes a negative pressure pump, and the negative pressure pump passes through a pipeline (which is provided with a pressure relief valve) and a water environment DNA with a filter membrane. The sample filter cup is connected.

[0061] The test water samples were collected in Jiuxiang River (32.113681 north latitude, 118.945127 east longitude).

[0062] The filter adopts Jinteng water system filter membrane, the filter membrane pore size is 0.45μm, and the filter membrane diameter is 50mm.

[0063] In this embodiment, the steps of the dry storage method for the DNA samples in the water environment are as follows:

[0064] (1) Use the negative pressure filter device to filter the water sample. After the water sample is filtered, turn off the negative pressure pump and open the pressure relief valve;

[0065] (2) After the pressure is zero, close the pressure relief valve, ...

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Abstract

The invention discloses a collection and normal-temperature drying storage method of water environment DNA samples, and belongs to the technical field of environment detection. The normal-temperaturedrying storage method of the DNA sample in the water environment comprises the following steps: collecting the sample in a water body by utilizing a filter membrane; after collection is completed, sequentially using isopropanol and ethyl alcohol to treat the filter membrane under a negative pressure adsorption condition; and finally, further air-drying the filter membrane, and then performing sealing storage, wherein the use amount of the ethyl alcohol is 1.2-1.5 times that of the isopropanol. According to the method, the storage time of the environmental sample can be effectively prolonged, the storage condition of the environmental water sample is reduced, and long-term storage of the environment DNA samples in the filter membrane under a normal temperature condition is realized, so thatcost is remarkably reduced, time efficiency is improved, and subsequent DNA monitoring is not interfered.

Description

technical field [0001] The invention belongs to the technical field of environmental detection, and more specifically relates to a method for collecting and storing water environment DNA samples. Background technique [0002] As we all know, the investigation of species richness is based on the concentration of environmental DNA, which is affected by the release and degradation process of DNA. At the same time, the extraction method of environmental DNA and the volume of water samples will also affect the concentration of environmental DNA. The eDNA metabarcoding technology mainly detects environmental DNA based on the acquisition of plankton (including plants, animals, microorganisms, and organisms of different sizes) in the receiving water and suspended particles in water samples. Therefore, if In order to truly reflect the DNA in the corresponding water environment, it is necessary to ensure the authenticity and validity of the collected samples. In order to minimize the...

Claims

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Application Information

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IPC IPC(8): C12Q1/24
CPCC12Q1/24
Inventor 杨江华张效伟
Owner 南京易基诺环保科技有限公司
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