Method for extracting fish genome DNA

A genome and fish technology, applied in the field of genomic DNA extraction, can solve the problems of long time and heavy extraction workload, and achieve the effect of reducing reaction costs

Inactive Publication Date: 2011-02-09
HEILONGJIANG RIVER FISHERY RES INST CHINESE ACADEMY OF FISHERIES SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] The purpose of the present invention is to provide a method for extracting fish genome DNA in order to solve the problem of heavy workload and long time for extracting fish common PCR reaction templates

Method used

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  • Method for extracting fish genome DNA
  • Method for extracting fish genome DNA
  • Method for extracting fish genome DNA

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Experimental program
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specific Embodiment approach 1

[0019] Specific embodiment one: the method for extracting fish genomic DNA in this embodiment is realized according to the following steps: one, fish collection sample preparation; Two, fish genomic DNA extraction; Three, PCR amplification; Add the test tube containing the fish collection sample and submerge the fish collection sample, then place it in an environment of 95±2°C for 30-90 minutes, then ice-bath, add the neutralization reaction solution equal to the volume of the alkaline reaction solution and Shake for 2 to 3 minutes, then centrifuge and retain the centrifuged supernatant to obtain the fish genome DNA template; in step 2, the alkaline reaction solution is composed of disodium ethylenediaminetetraacetic acid solution and sodium hydroxide solution, and the ethyl alcohol in the alkaline reaction solution is The concentration of disodium diaminetetraacetic acid is 0.2mmol / L, the concentration of sodium hydroxide is 25mmol / L; The neutralization reaction solution in st...

specific Embodiment approach 2

[0023] Embodiment 2: The difference between this embodiment and Embodiment 1 is that the number of PCR amplification cycles in step 3 is 25-27. Other steps and parameters are the same as those in Embodiment 1.

specific Embodiment approach 3

[0024] Specific embodiment three: the difference between this embodiment and specific embodiment one or two is: Step 1 cuts off the soft fins with a fish thickness less than 1mm, takes tissue pieces less than 2.5mmx2.5mm and scrapes off the surface mucus, that is Obtain fish collection samples; or take freshly hatched unopened fry with body height and body thickness less than 2mm, unhatched fertilized eggs with egg diameter of 0.8-1.2mm or animal poles with large egg diameter fertilized eggs as fish collection samples. Other steps and parameters are the same as those in Embodiment 1 or 2.

[0025] For fish that have not formed hard fins, the soft fins can be directly taken, and for fish with hard fins, the soft fins of the thinner and tender parts, such as the tip of the caudal fin, can be directly taken.

[0026] The present embodiment adopts soft fins to be broken as far as possible after removing the surface layer of mucus.

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Abstract

The invention discloses a method for extracting fish genome DNA, relates to a method for extracting genome DNA, and solves the problems of high workload and long time during extraction by a conventional common fish PCR reaction template. The method comprises the following steps of: 1, preparing acquired fish samples; 2, extracting the fish genome DNA; and 3, performing PCR amplification. The method can be used for extracting the fish genome DNA.

Description

technical field [0001] The invention relates to a method for extracting genome DNA. Background technique [0002] Polymerase Chain Reaction (PCR) is an in vitro nucleic acid amplification technology developed in the mid-1980s. It has the characteristics of specificity, sensitivity, speed, simplicity, good repeatability and easy automation. It has become a field of biological research. Indispensable research method. In the biological research of aquatic fish, the PCR method is the first method for nucleic acid detection. [0003] Currently, proteinase K is mainly used for genomic DNA extraction. First, digest the sample with proteinase K, then use phenol / chloroform to extract 2 to 3 times to remove impurity proteins. If the concentration of salt ions is high, dialyze it, then add 2 times the volume of absolute ethanol for precipitation, and wash with 75% alcohol. After drying, add 1 / 10TE to dissolve. This process takes a long time and requires a lot of work. Especially fo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/10
Inventor 贾智英石连玉
Owner HEILONGJIANG RIVER FISHERY RES INST CHINESE ACADEMY OF FISHERIES SCI
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