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Water environment DNA collection and preservation card and preparation method thereof

A technology for preserving card and water environment, which is applied in the direction of biochemical equipment and methods, microbial measurement/inspection, etc. It can solve the problems of high storage and transportation conditions, limited storage time, and increased costs, so as to avoid high costs, The effect of long storage time and cost saving

Pending Publication Date: 2020-01-03
李润成
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to provide a water environment DNA collection and storage card and its production method, to solve the problem that in the current conventional water environment DNA research proposed in the above background technology, because the collection site does not have DNA separation and purification conditions, it is necessary to carry water samples Back to the laboratory for environmental sample processing and DNA extraction, high storage and transportation conditions, increased costs, and limited storage time

Method used

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  • Water environment DNA collection and preservation card and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1: Preparation of DNA collection and preservation card in water environment 1

[0025] 1) Dissolve 3g Tris alkali and 3.72g EDTANa with 300ml deionized water 2 ﹒ H 2 0, 25g SDS, 118g guanidine isothiocyanate, 1.9mg thiourea, 1% NP40 and 4.383g NaCl, adjust the pH to 8.0 with concentrated hydrochloric acid, add deionized water to make the volume up to 500ml;

[0026] 2) Cut a certain size of nitrocellulose membrane with a thickness of about 0.3mm and put it into a certain volume of the above solution (according to 50μL solution / cm 2 The ratio of adsorbent material), placed in a closed container to prevent external DNA contamination, shake at 100rpm for 30 minutes, until the solution is completely absorbed.

[0027] 3) Carefully take out the card that has absorbed the salt solution, bake it at 80°C for 1 hour, and dry it sufficiently.

Embodiment 2

[0028] Example 2: Making a DNA collection and preservation card for water environment 2

[0029] 1) Dissolve 3g Tris alkali and 1.86g EDTANa in 300ml deionized water 2 ﹒ H 2 0, 10g SDS, 59g guanidine isothiocyanate, 0.95mg thiourea, 1% NP40 and 2.191g NaCl, adjust the pH to 8.0 with concentrated hydrochloric acid, and add deionized water to make the volume up to 500ml;

[0030] 2) Cut a certain size of nitrocellulose membrane with a thickness of about 0.3mm and put it into a certain volume of the above solution (according to 50μL solution / cm 2 The ratio of adsorbent material), placed in a closed container to prevent external DNA contamination, shake at 100rpm for 30 minutes, until the solution is completely absorbed.

[0031] 3) Carefully take out the card that has absorbed the salt solution, bake it at 80°C for 1 hour, and dry it sufficiently.

Embodiment 3

[0032] Example 3: Using DNA collection and preservation card to collect environmental samples

[0033] 1) Dissolve 3g Tris alkali and 3.72g EDTANa with 300ml deionized water 2 ﹒ H 2 0, 25g SDS, 118g guanidine isothiocyanate, 1.9mg thiourea, 1% NP40 and 4.383g NaCl, adjust the pH to 8.0 with concentrated hydrochloric acid, add deionized water to make the volume up to 500ml;

[0034] 2) Take 1L of water sample, filter with a filter membrane, collect 50-100mg of sediment on the filter membrane, add 50-100μL of the above solution, invert it up and down vigorously to mix it well, draw 50μL of lysate and add it carefully to Collect DNA on a sample card (approximately 2.5cm×2.5cm size card), spread it evenly, cover the entire card, and dry. Or pour the well-mixed lysate into a clean petri dish, and invade the DNA storage card with the size of 2.5cm×2.5cm into it, and then fully and uniformly adsorb the DNA of the sample in the water environment for a period of time and dry it.

[0035] 3)...

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Abstract

The invention discloses a water environment DNA collection and preservation card and a preparation method thereof, and relates to the technical field of biology. The problems that due to the lack of DNA separation and purification conditions at the collection site in existing conventional studies of water environment DNA, water samples need to be brought back to a laboratory for environmental sample processing and DNA extraction, storage and transportation condition requirements are high, the cost is increased, and the storage time is limited are aimed to be solved, the water environment DNA collection and preservation card includes a material with the ability to lyse cells and adsorb the DNA, the adsorption material is internally adsorbed with salinity, and the salinity includes a cell lysis agent, a nuclease inhibitor, a nucleic acid stabilizer and an antioxidant, wherein the cell lysis agent is one or more combinations of Tris hydrochloride, NP40 (ethyl phenyl polyethylene glycol),sodium chloride, guanidine isothiocyanate and SDS (sodium dodecyl sulfonate); and the nuclease inhibitor is a metal ion chelating agent, and the metal ion chelating agent is one of EDTA and an EDTA sodium salt.

Description

Technical field [0001] The invention relates to the field of biotechnology, and specifically relates to a DNA collection and preservation card for water environment and a manufacturing method thereof. Background technique [0002] Ecological environment and biodiversity research has always been the focus of attention in related fields. Environmental DNA usually refers to the sum of the DNA of various species obtained from environmental samples. It has attracted much attention because of its rapid identification of environmental biological species. Freshwater ecosystem is one of the important resources that people rely on for survival. In recent years, the deterioration of the ecological environment and the decrease of biodiversity have made the investigation of freshwater resources and the monitoring of aquatic species have become current research hotspots in the relevant fields of ecology. Compared with traditional species monitoring methods, environmental DNA monitoring has th...

Claims

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Application Information

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IPC IPC(8): C12Q1/6806
CPCC12Q1/6806
Inventor 李润成
Owner 李润成
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