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Lysis solution, extraction solution, lysis and extraction methods, kits and applications, PCR system

A technology for extracting liquid and lysing liquid, applied in the field of genetic engineering, can solve problems such as unfavorable fish research, and achieve the effects of being easy to master, simple and convenient to operate, and expanding the source of materials

Active Publication Date: 2019-05-28
CHINA JILIANG UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The existing animal tissue DNA extraction method is to remove the protein and fat in the tissue
Obviously, existing methods for extracting DNA from animal tissues are not conducive to fish research

Method used

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  • Lysis solution, extraction solution, lysis and extraction methods, kits and applications, PCR system
  • Lysis solution, extraction solution, lysis and extraction methods, kits and applications, PCR system
  • Lysis solution, extraction solution, lysis and extraction methods, kits and applications, PCR system

Examples

Experimental program
Comparison scheme
Effect test

Embodiment A1～A4

[0096] Embodiment A1~A4, embodiment B1~B4 and embodiment C1~C4

[0097] Extraction of DNA from small yellow croaker tissues. The small yellow croaker tissues used are scales, bones, muscles, and liver.

[0098] Embodiment A1~A4

[0099] Wash the small yellow croaker tissue with sterilized distilled water, take 25mg of the cleaned small yellow croaker tissue, cut it into pieces, put it into a 1.5mL centrifuge tube, add 400μL of lysate, shake for 20 seconds, keep it in a water bath at 55°C for 10min, then add 120μL The neutralization solution was shaken for 20 seconds, then centrifuged at 10,000g for 2 minutes, and the supernatant was taken to obtain a solution containing DNA.

[0100] The difference between the reaction conditions of Examples A1-A4 and Examples B1-B4 and Examples C1-C4 is only that the proportions of the respective reagents in the lysate and neutralization solution are different, see Table 1:

[0101] Table 1 The composition of reagents in lysate and neutral...

Embodiment D～ Embodiment S

[0119] The fish tissue DNA extraction solutions used in Examples D to S are the same as those in Examples C1 to C4, except for the type of small yellow croaker tissue, the amount of small yellow croaker tissue, lysate and neutralizing solution, and the water bath conditions.

[0120] Example

[0121] The DNA concentration of the DNA-containing solution prepared in Examples D to S was determined. The inventors found that the DNA concentration in the DNA-containing solution obtained from the treatment of muscle and liver was 50-200 ng / μL, and the concentration of DNA in the solution obtained from the treatment of fish fins contained The DNA concentration in the DNA solution is 20-100 ng / μL, and the DNA concentration in the DNA-containing solution obtained by treating the scales is 8-40 ng / μL. Moreover, in the above-mentioned solution containing DNA, A260 / A 280 The ratios are all between 1.2 and 1.6, indicating that the purity of the DNA obtained in the examples of the...

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Abstract

The invention discloses a pyrolysis solution, a DNA extracting solution, a pyrolysis and extraction method, a NDA extraction kit and application and a PCR (Polymerase Chain Reaction) system. The pyrolysis solution is used for splitting fish tissue to release DNA, and comprises ethylenediamine tetraacetic acid and / or ethylenediamine tetracetic acid disodium salt, sodium chloride, lauryl sodium sulfate, strong base and water, wherein the mole concentration of the ethylenediamine tetraacetic acid and / or the ethylenediamine tetracetic acid disodium salt is 2-10mM relative to the pyrolysis solution; the mole concentration of the sodium chloride is 1-10mM relative to the pyrolysis solution; the mass percentage of the lauryl sodium sulfate is 0.1-10% relative to the pyrolysis solution; the mole concentration of hydroxyl ions of the strong base is 100-300mM. Compared with the prior art, the pyrolysis solution disclosed by the invention is capable of providing a DNA template for fish tissue PCR amplification within relatively short time, reduction or loss of sample DNA can be avoided in the whole process, and moreover, operation is simple and convenient and easy to master.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to a lysate, a fish tissue DNA extraction solution, a lysis method, a method for extracting fish tissue DNA, a fish tissue DNA extraction kit and its application, and amplification of fish tissue DNA PCR system. Background technique [0002] The main methods for animal tissue DNA extraction include phenol-chloroform extraction after protease cleavage, isopropanol or isoamyl alcohol extraction, and SDS method, as well as cesium chloride precipitation, chelex-100 or Tween (Tween) for mitochondrial DNA extraction. ) method and urea extraction method, etc., the relatively simple and convenient commercial animal tissue DNA extraction kit is widely used at present. The genomic DNA obtained by these methods can generally remove the protein and fat in the original tissue sample, and can also remove phenols and other reagents through alcohol precipitation. The obtained DNA has a...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/10
CPCC12N15/1003C12Q1/686
Inventor 管峰薛超波赵进王萍亚
Owner CHINA JILIANG UNIV
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