Northeast China tiger hair DNA extracting method and microsatellite DNA label primer sequence

A technology of DNA labeling and primer sequence, which is applied in fermentation and other directions, and can solve the problems of unsuitable non-damaging DNA extraction

Inactive Publication Date: 2005-12-14
李迪强 +1
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  • Summary
  • Abstract
  • Description
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  • Application Information

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Problems solved by technology

Although these kits can quickly extract DNA, they are generally suitable for extraction materials such as blood samples or other damaged tissues, and are not suitable for non-damaging DNA extraction.

Method used

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Examples

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Embodiment Construction

[0014] The specific embodiment of the present invention is: the root of hair is put into the sterilized centrifuge tube, after washing with double distilled water, dehydrated ethanol soaks, and room temperature is dried; -HCl), ethylenediaminetetraacetic acid (EDTA), sodium chloride (NaCl), which also contains sodium dodecylbenzenesulfonate (SDS), dithiothreitol (DTT) and proteinase K, Obtain liquid A, keep it in a water bath at 37°C overnight; then extract liquid A twice: add an equal volume of saturated phenol to A for the first time, centrifuge at a medium speed for 10 minutes, and extract the supernatant to obtain solution B 1 , and repeat this step once to obtain solution B 2 ; the second time in B 2 Add an equal volume of chloroform, centrifuge at medium speed for 10 minutes, extract the supernatant to obtain solution C 1 , and then repeat this step once to obtain solution C 2 ; in C 2 Add 1 / 10 of the volume of sodium acetate solution and 2.5 times the volume of ice ...

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Abstract

The present invention is northeast China tiger hair DNA extracting method and microsatellite DNA label primer sequence. The hair DNA extracting process includes flushing the hair root inside sterilized centrifuge tube with double distilled water and alcohol and dry at room temperature; adding mixed solution of Tris-HCl, EDTA and NaCl and containing also SDS, DTT and proteinase K in 0.5 ml to obtain liquid A; twice extracting liquid A and separating for several times to obtain sample E; and obtaining the microsatellite DNA label primer sequence Pti002 of northeast China tiger. The obtained microsatellite DNA label primer sequence Pti002 is shown in the present invention.

Description

Technical field [0001] The invention relates to a method for extracting Amur tiger hair DNA and a microsatellite DNA marker primer sequence, which is particularly suitable for the analysis of Amur tiger DNA, and can also be used in the fields of Amur tiger population analysis, parent-child identification and the like. Background technique [0002] The Siberian tiger is listed as endangered by the International Union for the Conservation of Nature and Natural Resources (IUCN), and belongs to the first-level national key protected wild animals. The number of wild Siberian tigers in China is very small, only about 10. There is inbreeding in the breeding of Siberian tigers, and the mating method of one female and multiple males is often used. The offspring may have two or more fathers, which leads to confusion in blood relationship and unclear pedigree. Therefore, the Siberian tiger The genetic research of the Siberian tiger is of great significance to the protection of the Sibe...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P19/34
Inventor 李迪强张于光
Owner 李迪强
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