Method for extracting genome DNA from mammal solidification blood

A mammalian and genomic technology, applied in the field of animal genomic DNA extraction, can solve problems such as harmfulness to human body, long consumption time, cumbersome steps, etc., and achieve the effects of low cost, simple method and broad application prospect.

Inactive Publication Date: 2008-04-02
CHINA AGRI UNIV
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AI Technical Summary

Benefits of technology

This patented technology simplifies the way we recover genome DNA from cow's cloned blood during fetal gestation. Its advantages include quicker processing time without harmful chemical agents like benzene, which makes it ideal for studying bovine growth characteristics such as nutritional status, disease risk factors, etc., while also being useful for developing new methods that improve animal health through testing their effectiveness with regard to specific diseases associated therewith.

Problems solved by technology

This patented technical solution described by this patents relates to improving the process of isolating genetic material like nucleus acid or cellular components during research on animals' behavior without causing damage that could affect their health if taken out too early. Existing solutions require complicated procedures involving chemical agents with potentially hazardous effects.

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  • Method for extracting genome DNA from mammal solidification blood

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Embodiment 1

[0020] Example 1. Extraction of Genomic DNA and its Purity Detection from Holstein Dairy Cow Coagulated Blood

[0021] The formula of the lysate is: 150mM NaCl, 50mM Na 2 - EDTA and 2.0% (g / 100 mL lysate) SDS.

[0022] Genomic DNA is extracted from the coagulated blood of Holstein cows with the method of the present invention, specifically comprising the following steps:

[0023] 1) Take 400 μl frozen-thawed blood samples of Holstein cows and place them in Eppendorf centrifuge tubes, then use ophthalmic surgical scissors to cut the blood clots of mammals into fine particles; Washing with 70% ethanol and 0.9% NaCl solution in mass / volume percentage for 4 times.

[0024] 2) Cell lysis: Wash the broken coagulated blood with double distilled water to remove hemoglobin, then mix the broken coagulated blood with 400 μl lysate, add 10 μl proteinase K to the mixed solution to a final concentration of 360 mg / mL mixed solution, Then put the mixture in a water bath at 55°C (55-65°C is...

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Abstract

The invention discloses a method for extracting genomic DNA from coagulated blood of mammals, which includes the following steps that: 1), the coagulated blood of mammals is cut and rinsed with double distilled water to eliminate the hemachrome, blended with lysis buffer with the volume ratio of 3-5.5: 5, and added with proteinase K to obtain blended solution with the final concentration of 360-400mg/mL, which undergoes water bath under 55-65DEG C for 3-6 hours until no sticky gobbets exist in the solution; 2), the blended solution obtained by step 1) is mixed with 5.5-6.5M NaCL solution and chloroform with the volume ratio of 1: 0.5-0.9: 1.5-1.9, vibrated, and then centrifuged, with the supernate harvested and, 3) the supernate is mixed evenly in the volume ratio of 1: 0.8-1.2 with glacial ethanol with the volumetric concentration of 90-100 percent to obtain a white deposit as the genomic DNA. The invention can play an important role on the extraction of genomic DNA of mammals and the research on the functional genome of the cow and the other livestock, and have a great prospect.

Description

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Claims

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Application Information

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Owner CHINA AGRI UNIV
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