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Method for extracting genomic DNA from giant salamander skin mucus

A genome and body surface technology, applied in the field of bioengineering, can solve problems such as violation of protection, numerous operation steps, experimental operators and environmental hazards, and achieve the effect of convenient preservation

Inactive Publication Date: 2013-07-03
YANGTZE RIVER FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, some researches related to population genetics and molecular biology of giant salamanders have been carried out at home and abroad, but most of them use the muscle, liver, blood and other tissues of giant salamanders as materials to extract genomic DNA, causing damage to the body of giant salamanders, which violates the protection principle. The purpose is to cause a certain loss to the limited giant salamander resources
At the same time, the existing technology mostly uses the traditional phenol-chloroform method to extract the genome of the giant salamander, which has many steps, is time-consuming and laborious, and has certain hazards to experimental operators and the environment.

Method used

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  • Method for extracting genomic DNA from giant salamander skin mucus
  • Method for extracting genomic DNA from giant salamander skin mucus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] 1. Preparation of lysate

[0026] Mix 160 microliters of 1 mol / L sodium chloride (NaCl) aqueous solution, 40 microliters of 100 mmol / L tris-hydrochloride (Tris–HCl) aqueous solution with a pH value of 8.0, and 2 microliters of 100 mmol / L L with a pH of 8.0 ethylenediaminetetraacetic acid disodium (EDTANa 2 ) aqueous solution at room temperature, then add 160 microliters of 10% sodium dodecyl sulfate (SDS) aqueous solution and 20 microliters of proteinase K aqueous solution with a concentration of 10 mg / mL, and mix rapidly to prepare into the desired lysate.

[0027] 2. Preserve the samples: Collect giant salamander body surface mucus or body surface peeling skin from the body surface of giant salamander without trauma, put the collected mucus or body surface peeling skin in 90% ethanol for dehydration, and store it at room temperature. Press down and save for later use;

[0028] 3. Ethanol replacement: Take out the mucus or peeled skin obtained in step 2, put it in a...

example 2

[0035] 1. Preparation of lysate

[0036] Mix 160 microliters of 1 mol / L sodium chloride (NaCl) aqueous solution, 40 microliters of 100 mmol / L tris-hydrochloride (Tris–HCl) aqueous solution with a pH value of 8.0, and 2 microliters of 100 mmol / L L with a pH of 8.0 ethylenediaminetetraacetic acid disodium (EDTANa 2 ) aqueous solution at room temperature, then add 160 microliters of 10% sodium dodecyl sulfate (SDS) aqueous solution and 20 microliters of proteinase K aqueous solution with a concentration of 10 mg / mL, and mix rapidly to prepare into the desired lysate.

[0037] 2. Preserve the samples: Collect giant salamander body surface mucus or body surface peeling skin from the body surface of giant salamander without trauma, put the collected mucus or body surface peeling skin in 90% ethanol for dehydration, and store it at room temperature. Press down and save for later use;

[0038] 3. Ethanol replacement: Take out the mucus or peeled skin obtained in step 2, put it in a...

Embodiment 3

[0045] 1. Preparation of lysate

[0046] Mix 160 microliters of 1 mol / L sodium chloride (NaCl) aqueous solution, 40 microliters of 100 mmol / L tris-hydrochloride (Tris–HCl) aqueous solution with a pH value of 8.0, and 2 microliters of 100 mmol / L L with a pH of 8.0 ethylenediaminetetraacetic acid disodium (EDTANa 2 ) aqueous solution at room temperature, then add 160 microliters of 10% sodium dodecyl sulfate (SDS) aqueous solution and 20 microliters of proteinase K aqueous solution with a concentration of 10 mg / mL, and mix rapidly to prepare into the desired lysate.

[0047] 2. Preserve the samples: Collect giant salamander body surface mucus or body surface peeling skin from the body surface of giant salamander without trauma, put the collected mucus or body surface peeling skin in 90% ethanol for dehydration, and store it at room temperature. Press down and save for later use;

[0048] 3. Ethanol replacement: Take out the mucus or peeled skin obtained in step 2, put it in a...

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Abstract

The invention discloses a method for extracting genomic DNA from giant salamander skin mucus. The method comprises the following steps: 1, saving a sample; 2, replacing ethanol; 3, splitting a cell; 4, precipitating protein; 5 precipiating DNA; 6, washing the DNA; and 7, obtaining DNA, wherein required pyrolysis liquid can be obtained by mixing 160 microliter 1 mol / L nacl aqueous solution, 40 microliter 1 mol / L TRIS-HCI aquenous solution with pH value of 8.0, and 2 microliter 100mmol / L EDTANa2 aqueous solution with pH value of 8.0 uniformly at normal temperature, and then adding 160 microliter 10% of SDS aqueous solution and 20 microliter 10 mg / ml proteinase K aqueous solution to the mixture and quickly mixing the mixture. The method provided by the invention adopts giant salamander skin mucus or skin as an extracting material of the genomic DNA, so that the giant salamander organism will be protected without damage.

Description

technical field [0001] The invention relates to the field of bioengineering, in particular to a method for extracting genome DNA from body surface mucus of giant salamanders. Background technique [0002] giant salamander ( Andrias ) belongs to the family Cryptobranchidae, which belongs to the family Amphibians, and is the largest extant amphibian in the world. There are 2 genera and 3 species in the family Cryptobranchidae, of which there are 2 species in the genus Giant Salamander, namely the Chinese giant salamander ( Andrias davidianus Blanchard, 1871) and the Japanese giant salamander ( Andrias japonicus Temminck, 1836); Cryptobranchus only has Cryptobranchus ( Cryptobranshus alleganiensis Daudin, 1803) a species distributed in the northeastern United States, also known as the American giant salamander. Due to the continuous increase of human activities, such as the construction of reservoirs, deforestation, mining, and the use of chemical fertilizers and pesti...

Claims

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Application Information

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IPC IPC(8): C12N15/10
Inventor 张书环危起伟梁志强
Owner YANGTZE RIVER FISHERIES RES INST CHINESE ACAD OF FISHERY SCI
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