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Fish scale genome DNA (deoxyribonucleic acid) extraction method and special kit

A genome and kit technology, applied in the field of DNA preparation, can solve problems such as not seen, and achieve the effects of fast extraction, easy storage, and convenient material collection

Inactive Publication Date: 2012-07-25
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, there is no report on the DNA extraction kit of fish scales, especially there is no kit on the market that is cheap, easy to operate, and suitable for routine DNA extraction of a large number of samples in the analysis of population genetic diversity.

Method used

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  • Fish scale genome DNA (deoxyribonucleic acid) extraction method and special kit
  • Fish scale genome DNA (deoxyribonucleic acid) extraction method and special kit
  • Fish scale genome DNA (deoxyribonucleic acid) extraction method and special kit

Examples

Experimental program
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Effect test

Embodiment 1

[0042] The composition and preparation of embodiment 1 fish scale genomic DNA extraction kit and its use method

[0043] 1. Composition and preparation of fish scale genome DNA extraction kit

[0044] 1. Reagent composition:

[0045] Buffer-1; Buffer-2; Buffer-3; proteinase K; isopropanol; 70% ethanol; absolute ethanol; sterile double distilled water.

[0046] 2. Reagent preparation:

[0047] A) Buffer-1: 100mM Tris-HCl, 50mM EDTA, 1mM CaCl 2 It is composed of sodium lauryl sulfate with a mass percentage of 0.5%. When the temperature is low, Buffer-1 will precipitate. Before use, put it in a water bath at 56-65°C to dissolve, shake well before use;

[0048] B) Buffer-2: It consists of 6% sodium lauryl sulfate, 0.6M NaCl and 0.2M ethylenediaminetetraacetic acid. When the temperature is low, Buffer-2 will produce precipitation. Place it in Dissolve in a water bath at 56-65°C, shake well before use;

[0049] C) Buffer-3: Composed of 7.5M ammonium acetate solution, pre-cooled...

Embodiment 2

[0067] Application of Example 2 Fish Scale Genomic DNA Extraction Kit

[0068] Experiment 1. Genomic DNA extraction from naturally dried grass carp scales

[0069] According to the steps of Example 1, wherein the material used is grass carp scales after natural air-drying at room temperature for one week, and the extracted DNA is detected by agarose gel electrophoresis, and the results are shown in figure 1 .

[0070] Experiment 2. Genomic DNA extraction from fresh bream scales

[0071] According to the steps of Example 1, wherein the material used is fresh bream (commonly known as Wuchang fish) scales, and the extracted DNA is detected by agarose gel electrophoresis, and the results are shown in figure 2 .

[0072] Experiment 3. Genomic DNA extraction from the scales of bream after one week of ethanol treatment

[0073] According to the steps of Example 1, wherein the material is the bream scales treated with ethanol for one week, the extracted DNA is detected by agarose...

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Abstract

The invention belongs to the technical field of DNA (deoxyribonucleic acid) preparation, and in particular relates to a method for extracting genome DNA from a fish scale and a special kit thereof. The kit provided by the invention comprises the following reagents: a) Buffer-1, b) Buffer-2, c) Buffer-3 and d) protease K and other reagents. The kit provided by the invention is simple and convenient to operate, is low in cost and is applicable to conventional DNA extraction of a large amount of samples during population genetic diversity analysis. The method provided by the invention is fast in extraction speed, and sample processing and whole sample DNA extraction can be completed within two hours; material is convenient to draw, a sample is easy to store, and the normal growth of fishes can not be influenced generally; and extracted DNA can be directly applied to PCR (polymerase chain reaction) amplification and related applications. The invention also discloses a method and steps for extracting the genome DNA from the fish scale.

Description

technical field [0001] The invention belongs to the technical field of DNA preparation, and in particular relates to a method for extracting genome DNA from fish scales and a special kit. Background technique [0002] Identification of fish germplasm and diversity assessment at the nucleic acid molecular level, as well as discussing their phylogenetic relationship and phylogenetic evolution, are currently effective means for fish genetics and breeding, resource assessment, and origin and evolution research. Genomic DNA Extraction is the basis of these works. [0003] The technology of convenient sampling and easy operation, high efficiency, low cost and adaptability to routine DNA extraction of a large number of samples and the corresponding kit preparation and application methods have always been the pursuit goals of researchers, and they are also the research personnel engaged in fish genetics and breeding. The needs of grass-roots science and technology extension personn...

Claims

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Application Information

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IPC IPC(8): C12N15/10
Inventor 王卫民李艳和高泽霞
Owner HUAZHONG AGRI UNIV
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