Cockle shellfish mitochondria COI (cytochrome c oxidase I) gene amplification primer

Abstract

The invention uses a universal primer for amplification to obtain 66 cardiidae mtCOI (cytochrome c oxidase I) sequences, uses a CLUSTALW software to compare and analyze, uses primer design software primer Premier5 to design 5 pairs of primers in a determined area, uses 216 cardiidae individuals to repeatedly conduct FCR reaction tests in a 10mu m system, finally is detected through agarose gel electrophoresis, and screens out ctockle shellfish mitochondria COI gene sequence efficient amplification primer. The cockle shellfish mitochondria COI gene amplification primer is characterized in that NGF5'-ACAAATCATAAAGATATTGG-3'NGR5'-TTCAGGGTGTCCGAAGAATCA-3' adopts the cockle shellfish mitochondria COI primer, objective sections of different species of cockle shellfish are effectively amplified, requirements of genetic diversity survey of cockles can be met, simultaneously reliable molecular technique instruments for germplasm resource protection and breeding industry development are provided.

Description

technical field [0001] The invention relates to a cockle shellfish mitochondrial COI [the cytochrome c oxidase I] gene amplification primer. Background technique [0002] The family Cardiidae belongs to the class Bivalvia, subclass Heterodonta, order Veneroida, and it is one of the largest and oldest bivalve shellfish. There are about 250 species of cockle shellfish in the world, and about 50 species have been found in my country. The diameter of the small one is about 1 cm, and the diameter of the large one can reach 15 cm (such as the smooth giant cockle in California). Cockles are distributed worldwide, often inhabiting the shallow seas of the Indian Ocean-Western Pacific Ocean, and are distributed in Singapore, Japan, Australia, Malaysia, Indonesia, mainland China, Taiwan and the South China Sea Islands. Many species in the cockle family are delicious in meat, rich in nutrition, have high commercial value, and are important breeding objects (such as sliding top thin-she...

AI Technical Summary

Problems solved by technology

[0004] However, the mutation rate of shellfish mtCOI sequence is significantly higher than that of other groups of animals, and the mtCOI sequence variation among different mollusk species, even related species or even different groups within a species, is very large, which led to Folmer et al. 1994 The amplification efficiency of the COI universal primers LCO1490 and HCO2198 designed in 2009 is extremely low in this animal group, and some species cannot be amplified at all, which hinders the use of molecular techniques