Single nucleotide polymorphic locus of cattle WNT10B gene, and detection method for single nucleotide polymorphic locus
A single nucleotide polymorphism and detection method technology, applied in the field of molecular genetics, can solve problems such as expensive and cumbersome operations
Inactive Publication Date: 2012-08-29
XUZHOU NORMAL UNIVERSITY
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The RFLP-PCR method not only has the accuracy of DNA sequencing, but also overcomes the shortcomings of expensive, cumbersome operations, and false positives, and there are no special requirements for the sequence sites detected
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[0047] In order to facilitate the understanding of the technical solution of the present invention, the present invention will be further described below in conjunction with specific examples.
[0048] The following is through the collection of cattle samples and genomic DNA extraction, detection, purification and concentration analysis, the PCR amplification of the first intron and the second, fourth and fifth exons of the cattle WNT10B gene, the first intron of the cattle WNT10B gene and the The PCR-RFLP analysis embodiment of the 2nd, 4th, 5th exon will further illustrate the technology of the present invention and its effect. What has been described is by way of explanation, not limitation, of the invention.
[0049] 1. Amplification of partial DNA sequence of cattle WNT10B gene and detection of its polymorphism
[0050] 1. Collection and processing of cattle blood samples
[0051] Take 10 mL of cattle blood sample, add 500 μL of 0.5 mol / L EDTA to anticoagulate, slowly i...
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The invention discloses a method for quickly detecting the single nucleotide polymorphism of a cattle WNT10B gene. The method comprises the following steps: performing polymerase chain reaction (PCR) amplification on the cattle WNT10B gene by using the whole genome DNA containing the WNT10B gene of the cattle to be detected as a template and primer pairs P1 and P3 as primers; respectively digesting the PCR products amplified by the primer pairs P1 and P3 by using restriction endonuclease NaeI and ApaI, and performing agarose gel electrophoresis on the amplification segment after enzyme digestion; and identifying the single nucleotide polymorphism of the 220th and the 3980th site of the cattle WNT10B gene according to the result of the agarose gel electrophoresis. The method is used for screening and detecting the genetic marker closely related to the growth and development traits of the cattle at the DNA level, so that the method is applied to assisted selection and molecular breeding of the cattle and the breeding speed of the excellent cattle species is increased.
Description
technical field [0001] The invention belongs to the field of molecular genetics, relates to the detection of gene single nucleotide polymorphism (SNP), in particular to a single nucleotide polymorphism for detecting intron 1 and exon 2, 4, 5 of cattle WNT10B gene. state-of-the-art detection method. Background technique [0002] Single nucleotide polymorphism (SNP) refers to the polymorphism caused by the substitution of a single nucleotide (A / T / C / G) in the genomic DNA sequence. Therefore, commonly referred to as SNPs include base substitutions, insertions, deletions, and changes in the copy number of repeated sequences. A SNP indicates that there is a nucleotide change at a certain position in the genome, which is mainly caused by the conversion or transversion of a single base; SNPs with conversion mutations account for about 2 / 3, and other SNPs are in similar level. The cytosine of the CpG dinucleotide is the most mutated site in the genome, most of which are methylated...
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Inventor 陈宏赵静张春雷房兴堂
Owner XUZHOU NORMAL UNIVERSITY