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PCR (Polymerase Chain Reaction) method for sex appraisal of Nipponia nippon

A crested ibis and gender technology, applied in the field of molecular biology, can solve the problems of misjudgment, lack of positive control, and rarely used, etc., and achieve the effect of strong repeatability, efficiency and accuracy

Inactive Publication Date: 2012-08-22
BEIJING FORESTRY UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the lack of positive controls in this method, it is easy to cause misjudgment (the result "no band" caused by experimental errors is misjudged as male), and it is rarely used in actual protection and research.
There is no report on the sex identification of crested ibis using CHD gene

Method used

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  • PCR (Polymerase Chain Reaction) method for sex appraisal of Nipponia nippon
  • PCR (Polymerase Chain Reaction) method for sex appraisal of Nipponia nippon
  • PCR (Polymerase Chain Reaction) method for sex appraisal of Nipponia nippon

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Embodiment 1

[0028] Embodiment 1, PCR identification crested ibis gender

[0029] The sexes of 33 crested ibises (Nipponia Nippon, Ding CQ. 2010. Crested Ibis. Chinese Birds. 1(2): 156-162, available from Beijing Forestry University) in Hanzhong, Shaanxi Province were identified. 1 body feather.

[0030] 1. Genomic DNA extraction of crested ibis feathers:

[0031] 1) Take a sterilized 2.0ml centrifuge tube and place it in a test tube rack, numbered 1-33;

[0032] 2) Use sterilized tweezers to take a clean feather from each crested ibis, use ophthalmic scissors to cut a 1cm length of the feather pith, and cut it into pieces as much as possible;

[0033] 3) Add 700μl STE solution (10mL Tris-HCl (1M; pH8.0), 2mL EDTA (0.5M; pH 8.0), 100mL 1M NaCl aqueous solution, distilled water to 1L to each centrifuge tube , autoclaved), 5 μl 10% SDS (mass percentage content) and 15 μl proteinase K (20 mg / mL), placed in a constant temperature water bath at 60 r / m for overnight digestion (18-20 hours); ...

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Abstract

The invention discloses a PCR (Polymerase Chain Reaction) method for the sex appraisal of Nipponia nippon, and provides a primer pair for detecting the sex of a bird. In the primer pair for detecting the sex of a bird, one primer is the nucleotide disclosed as sequence 1 in the sequence list, and the other primer is the nucleotide disclosed as sequence 2 in the sequence list. Experiments providedby the invention prove that the sex of Nipponia nippon can be conveniently and accurately appraised based on different banding patterns produced by PCR amplification by using cross-intron primers designed on the basis of the length difference between Z and W chromosomes of CHD gene introns. The technology is of far reaching importance in the sex ratio monitoring of wild young Nipponia nippon, theconstruction and reintroduction of the scientific pairing system of breeding population as well as the selection and reasonable collocation of released individuals.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a PCR method for gender identification of crested ibis. Background technique [0002] The CHD gene of birds, namely the chromo-helicase-DNA binding gene, is located on the sex chromosomes. The sex chromosomes of birds are ZW type, males are ZZ, and females are ZW. The CHD gene has two homologous copies CHD-W and CHD-Z in most non-ratite birds, where CHD-W is W chromosome linkage and CHD-Z is Z chromosome linkage. Because the exons of the CHD gene are highly conserved, while the introns are more mutated between the sexes, it has become the most important molecular marker for sex identification of non-ratite birds. [0003] Gender is one of the most important characteristics of birds. Male and female birds often have differences in physiology and behavior. The sex identification of birds, especially the early identification of sex, is of great significance to the preven...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/68
Inventor 丁长青和雪莲韩建林
Owner BEIJING FORESTRY UNIVERSITY
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