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Extraction method of cubilose whole genome DNA

An extraction method and a genome-wide technology, applied in the field of biochemistry, can solve the problems of low DNA purity and difficult experiments, achieve good reproducibility, stable results, and reduce co-precipitation of glycoproteins and DNA

Inactive Publication Date: 2009-05-06
GUANGZHOU UNIVERSITY OF CHINESE MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] However, the main component of bird's nest saliva is sialoglycoprotein, and its molecular structure contains peptide chains and sugar chains, so it must have both the physical and chemical properties of proteins and polysaccharides. When extracting DNA, a large amount of sialoglycoproteins are easy to be combined with DNA Precipitation, therefore, no matter which of the above extraction methods is used, the purity of the resulting DNA is not high, making downstream experiments difficult

Method used

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  • Extraction method of cubilose whole genome DNA
  • Extraction method of cubilose whole genome DNA

Examples

Experimental program
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Effect test

example 1

[0032] 1. Material: Baiyan bird's nest, purchased from Singapore Jinxing Company.

[0033] 2. Method:

[0034] Take the dry product of Baiyan bird's nest and grind it into a 120-mesh fine powder, weigh 0.1200g, add 1.2ml of SDS buffer, bathe in water at 65°C for 2 hours, and shake gently 4 times in the first half hour. Centrifuge at 12000×g for 5 min at 4°C to take the supernatant. An equal volume of chloroform-isoamyl alcohol mixture was added to the obtained supernatant, and after mixing evenly, the supernatant was collected by centrifugation at 12000×g for 5 min at 4°C. Add 0.1 volume of CTAB buffer solution to the obtained supernatant at room temperature (20°C), mix evenly, place it at room temperature for 15 minutes, then add chloroform-isoamyl alcohol mixed solution equal to the volume of the mixed solution, mix well, and place at 4°C at 12000 Centrifuge at × g for 5 min to take the supernatant. Add 0.7 volume of isopropanol at -20°C to the obtained supernatant, mix w...

example 2

[0040] 1. Material: Xueyan bird's nest, purchased from Singapore Jinxing Company

[0041] 2. Method:

[0042]Take the dried bird's nest product of Xueyan and grind it into a 120-mesh fine powder, weigh 0.1200g, add 1.2ml of SDS buffer, bathe in water at 65°C for 2 hours, and shake gently 4 times in the first half hour. Centrifuge at 12000×g for 5 min at 4°C to take the supernatant. An equal volume of chloroform-isoamyl alcohol mixture was added to the obtained supernatant, and after mixing evenly, the supernatant was collected by centrifugation at 12000×g for 5 min at 4°C. Add 0.1 volume of CTAB buffer solution to the obtained supernatant at room temperature (20°C), mix evenly, place it at room temperature for 15 minutes, then add chloroform-isoamyl alcohol mixed solution equal to the volume of the mixed solution, mix well, and place at 4°C at 12000 Centrifuge at × g for 5 min to take the supernatant. Add 0.7 volume of isopropanol at -20°C to the obtained supernatant, mix w...

example 3

[0048] 1. Material: Swiftlet bird’s nest, purchased from Singapore Jinxing Company

[0049] 2. Method:

[0050] Take the dried swiftlet nest and grind it into a 120-mesh fine powder, weigh 0.1200g, add 1.2ml of SDS buffer, bathe in water at 65°C for 2 hours, and shake gently 4 times in the first half hour. Centrifuge at 12000×g for 5 min at 4°C to take the supernatant. An equal volume of chloroform-isoamyl alcohol mixture was added to the obtained supernatant, and after mixing evenly, the supernatant was collected by centrifugation at 12000×g for 5 min at 4°C. Add 0.1 volume of CTAB buffer solution to the obtained supernatant at room temperature (20°C), mix evenly, place it at room temperature for 15 minutes, then add chloroform-isoamyl alcohol mixed solution equal to the volume of the mixed solution, mix well, and place at 4°C at 12000 Centrifuge at × g for 5 min to take the supernatant. Add 0.7 volume of isopropanol at -20°C to the obtained supernatant, mix well and immed...

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Abstract

The invention provides a method for extracting genomic DNA from swallow nests, which comprises the following steps: firstly, lysing cells in the swallow nest by use of SDS and DTT, ,and simultaneously removing most of glycosidoprotein through high-concentration NaCl, so as to reduce the interference of glycosidoprotein to DNA in the purification process; secondly, further removing the glycosidoprotein by use of combining chloroform-isoamyl aleohl with CTAB; and finally, depositing DNA by isopropanol at a low temperature in a short time, which can reduce coprecipitation of glycosidoprotein and DNA to the utmost extent. The method has the advantages of stable result, good reproduction quality, simple operation, low cost, and no need of special reagent and apparatus; moreover, the method has high quality of the obtained genomic DNA which can meet the requirement of the PCR reaction, and can realize identification of the variety of the swallow nests.

Description

technical field [0001] The invention relates to the field of biochemistry, in particular to a method for extracting deoxynucleic acid. Background technique [0002] The common bird's nest is a jelly-dried saliva secreted by various birds of the family Apodidae (Aerodramus or Collocalia) or a mixture of saliva and their down feathers. The dried saliva secreted by some birds of the genus Swift (Apus) when they build nests, such as the bird's nest produced in Huaiji County, Guangdong Province, China (also called Longya Bird's Nest in some places). Because bird's nest is a valuable tonic, the high market price has led unscrupulous traders to confuse the real ones with substandard ones. Many fake and counterfeit products are difficult to distinguish from the appearance, and the DNA in the bird's nest mainly comes from oral epithelial cells shed in saliva and down feathers. Hair follicle cells cannot be forged. Therefore, molecular detection methods are particularly important in ...

Claims

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Application Information

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IPC IPC(8): C07H21/04C12N15/10
Inventor 赖小平林洁茹黄松王培训周华陈建南蒋东旭候雁冼小敏
Owner GUANGZHOU UNIVERSITY OF CHINESE MEDICINE
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