Method for extracting genome DNA (deoxyribonucleic acid) of rhinopithecus brelichi through feces sampling

A technology of the golden snub-nosed monkey and the genome is applied in the field of feces sampling to extract the genomic DNA of the golden snub-nosed monkey, which can solve problems such as difficult population genetic analysis, and achieve the effects of high purity, simple operation and low reagent toxicity.

Inactive Publication Date: 2015-04-01
ZUNYI MEDICAL UNIVERSITY
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Problems solved by technology

Even in species with a large number of individuals, the collection of blood samples can only be limited to a small number of individuals, and it is difficult to carry out larg

Method used

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[0023] The present invention will be described in further detail below:

[0024] A method for fecal sampling and extraction of genomic DNA of Qianjin snub-nosed monkey includes the following steps:

[0025] Step 1: Take 2.5 g of Guizhou golden monkey feces frozen at -20°C for easy sterilization, add 30ml TE to dissolve, stir well and let stand for 2 min;

[0026] Step 2: Take 1 mL of the supernatant into a 1.5 mL Eppendorf tube, make 10 tubes, 4℃, 500 r / min, centrifuge for 5 min;

[0027] Step 3: Take the supernatant in another Eppendorf tube and centrifuge at 1500 r / min at 4°C for 5 minutes to obtain exfoliated cells from the intestinal wall;

[0028] Step 4: Add 0.5 mL of acetone pre-cooled at -20°C, wash the precipitate, 4°C, 1500 r / min, centrifuge for 5 min, pour off the acetone, repeat the washing 3 times, and dry;

[0029] Step 5: Add 600 μL of TE and 40 μL of lysozyme at a concentration of 50 mg / mL and incubate at 37°C for 30 min; centrifuge at 1 500 r / min at 4°C for 5 min, and di...

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Abstract

A method for extracting genome DNA (deoxyribonucleic acid) of rhinopithecus brelichi through feces sampling comprises steps as follows: cryopreserved feces of rhinopithecus brelichi is taken and placed in a sterilized beaker, TE is added, and the feces is dissolved, stirred and left to standing; a supernatant is taken and placed in ten Eppendorf tubes and is centrifuged; a supernatant is taken and placed in another Eppendorf tube and is centrifuged to obtain intestinal wall exfoliative cells; precooled acetone is added, precipitate is washed, centrifugation is performed, acetone is poured out, and washing is repeatedly performed for three times, and a product is dried; TE and lysozyme are added, and heat preservation is performed at the temperature of 37 DEG C; centrifugation is performed, supernatant is abandoned; lysate and proteinase K are added to the precipitate, and the precipitate is subjected to warm bath for longer than 3 h at the temperature of 55 DEG C; RNAse (ribonuclease) is added, and heat preservation is performed at the temperature of 37 DEG C; the Eppendorf tubes are placed in boiling water bath at the temperature of 94 DEG C; centrifugation is performed, supernatant is taken, double volumes of absolute ethyl alcohol is added, and the mixture is uniformly mixed and placed for longer than 30 min at the temperature of 20 DEG C; and centrifugation is performed, precipitate is washed by precooled 70% ethyl alcohol twice, then centrifugation is performed, and TE is added to dissolve DNA after the precipitate is dried, and storage is performed at the temperature of subzero 20 DEG C.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a method for extracting genome DNA of Guizhou golden monkey by feces sampling. Background technique [0002] Guizhou golden monkey ( Rhinopithecus brelichi ) belonging to the primate order ( Primates ) Monkey family ( Cercopithecidae ) snub-nosed monkey ( Rhinopithecus ), which is the prototype of the legendary Monkey King Sun Wukong. It has a lovely appearance and is of considerable ornamental value. Among the three species of golden monkeys endemic to my country, the Guizhou snub-nosed monkey has the least number and the narrowest habitat. It is a rare protected animal and is known as the "only child in the world". Within the scope of square kilometers, it is even rarer than the national treasure giant panda. As early as 1989, it was listed as a first-class key protected animal in my country; in 1996, it was included in the Red Data Book of Endangered Animals (E-level) [1] . ...

Claims

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Application Information

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IPC IPC(8): C12N15/10
Inventor 张志敏肖代敏李学英潘有福钱刚
Owner ZUNYI MEDICAL UNIVERSITY
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