Amplification primer for grouper catostomidae mitochondrial genome complete sequence and design method thereof

Abstract

The invention discloses an amplification primer for a grouper catostomidae mitochondrial genome complete sequence and a design method thereof, relates to groupers, and particularly relates to 16 pairs of amplification primers mainly used for specifically amplifying most grouper catostomidae mitochondrial genome complete sequences and a design method thereof. The invention provides an amplification primer for a grouper catostomidae mitochondrial genome complete sequence, a design method thereof and application thereof. The amplification primer for the grouper catostomidae mitochondrial genome complete sequence consists of 16 pairs of single-chain oligonucleotides, wherein each pair consists of one light-chain primer and one heavy-chain primer. The amplification primer for the grouper catostomidae mitochondrial genome complete sequence can be used in the aspects of variety authentication, geographical population authentication, germplasm resource evaluation and the like.

Description

technical field [0001] The invention relates to grouper, in particular to 16 pairs of amplification primers mainly used for specifically amplifying the complete mitochondrial genome sequence of most grouper subfamily fishes and a design method thereof. Background technique [0002] The mitochondrial genome consists of 37 coding genes and a major non-coding region (control region), and the evolution rates of different sequence regions are different. Because animal mitochondrial DNA (mtDNA) has the characteristics of simple structure, strict maternal inheritance, almost no recombination, fast evolution rate and differences in evolution rate in different regions, it is widely used in species identification, phylogenetic evolution and population genetics research. Very broad. [0003] At present, it is a very convenient and accurate method to identify fish with similar appearance through sequence differences. Among them, the study of mitochondrial sequence differences, such as ...

AI Technical Summary

Problems solved by technology

However, due to the lack of primers suitable for the amplification and sequencing of the complete mitochondrial genome sequence, the determination of the complete mitochondrial genome sequence of the grouper is progressing slowly

Meyer (4, MeyerA (1993) Evolution of mitochondrial DNA in fishes.In: Biochemistry and Molecular Biology of Fishes (eds.Hochanchka PW, MommsenTP), vol.2, 1-38, Elsevier, NewYork) once announced in 1993 that fish General amplification primers for mitochondrial genomes, but grouper mitochondrial genome sequences are relatively specialized, and it is difficult to effectively amplify them with general amplification primers

So far, only Zhu et al. (5, Zhu ZY, Yue GH (2008) The complete mitochondrial genome of red grouper Plectropomus leopardus and its applications in identification of grouper species. Aquaculture, 276, 44-49) successfully utilized Meyer's universal amplification primers Amplify the complete mitochondrial genome sequence of a grouper fish, and these primers have high requirements on the quality of DNA templates, long-chain PCR amplification and step-by-step sequencing, which are difficult and cumbersome