Molecular marking method of using neuroendocrine factor genes to select kidding characters

Abstract

The invention discloses a molecular marking method of using neuroendocrine factor genes to select kidding characters. The method comprises the following steps of: using a goat genomic DNA sequence as a template; amplifying Kisspeptin gene intron 2 by using a primer P1 under the PCR condition in the presence of TaqDNA polymerase, buffering environment, Mg2+, dNTPs, and then judging the size of the target fragment by agarose gel electrophoresis; detecting the PCR application product of the primer P1 by polyacrylamide gel electrophoresis, finding that the amplification SNPs of the primer P1 has one-base mutation and 8bp-base deletion, and then carrying out genotyping and gene frequency analysis on the amplification product of the primer P1, and the correlation analysis with the kidding characters of Guanzhong diary goat and Xinong Saanen dairy goat. Shown by experiments, the SNPs of the Kisspeptin gene intron 2 detected by the primer P1can be used as molecular marker of the selection of kidding characters.

Description

technical field [0001] The invention belongs to the field of molecular genetics, and specifically relates to a molecular marker method for selecting lambing traits of goats by using neuroendocrine factor genes. The results showed that the SNPs detected by primer P1 in intron 2 of kisspeptin gene could be used as molecular markers for selection of lambing traits in goats. Background technique [0002] With the rapid development of molecular biology technology, especially the advent of Polymerase Chain Reaction (Polymerase Chain Reaction) technology and the improvement of electrophoresis technology, various molecular genetic marker technologies have emerged. Breeding of varieties provides new ways and methods. In particular, the combination of molecular marker technology and conventional breeding technology has given birth to a brand-new selection method, that is, marker-assisted selection (Marker assistant selection). It can overcome the environmental deviation and sampling...

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Problems solved by technology

Among these SNP detection techniques, DNA sequence determination is the most accurate SNP detection method, but its detection cost is extremely expensive, and large-scale instruments such as DNA sequencers are required. At the same time, very skilled technicians and Experience, so DNA sequence determination is not an ideal SNP detection method for actual production; of course, the combination of PCR-SSCP and DNA sequencing can be used to detect SNPs can appropriately reduce the detection cost, but the experimental process of PCR-SSCP is relatively long , the operation is cumbersome, and there are false negative problems in the experimental process, so it is not an ideal SNP detection method; as a new SNP detection method, the AS-PCR method has very broad prospects in the future application field, but , this method needs to design special primers, and it can only target specific gene loci. At the same time, there is a probability of false detection in the detection process. Therefore, it is not generally applicable at present; Ligation reaction technology to detect SNP sites requires detection platforms such as plate reader, gene chip, microsphere array technology and mass spectrometer, which is not very practical for general molecular laboratories

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