Inverse PCR amplification of duck circovirus gene

Abstract

The invention discloses inverse PCR amplification of a duck circovirus gene. The inverse PCR amplification is characterized in that the inverse PCR amplification includes subjecting a PCR product to electrophoresis on agarose gel having a concentration of 2% and cutting a target fragment under a fluorescent lamp; recovering and purifying; dipping into an LB solution and performing blue / white selection to screen again; subjecting selected bacterial colonies to plasmid extraction, performing PCR again and performing restriction enzyme digestion identification; performing pre-degeneration in an environment having a temperature of 95-100 DEG C for 3-4 min; and then repeating the above step every 3 min for 25 times; agarose gel electrophoresis detection is performed again after the step is repeated for 25 times; and ice bath is performed for 3 min before electrophoresis detection. Beneficial effects of the inverse PCR amplification are that successful extraction of DNA in the duck circovirus genome, high purity, simple and convenient operation, low cost, strong repeatability and capability of meeting experiment standards are achieved.

Description

technical field [0001] The invention relates to an experimental method, in particular to the reverse amplification of duck circovirus gene PCR. Background technique [0002] DuCV belongs to the family Circoviridae and the genus Circovirus. It has no envelope, a single-stranded circular DNA structure, and icosahedral symmetry. Its size under the electron microscope is similar to that of other circoviruses, with a diameter of 15 to 16 nm. Since it is not yet possible to isolate and culture it, the research on its etiological characteristics is quite limited. The common feature of circoviruses is that they cause host immunosuppression and lead to secondary infection or double and multiple infections, especially chicken infectious anemia virus (CIAV) and porcine circovirus type 2 (PCV2) in recent years. The industry has caused huge economic losses, which has attracted great attention to circovirus all over the world. Avian circovirus infection is characterized by clinical sym...

AI Technical Summary

Problems solved by technology

Since isolation and culture cannot be carried out at present, the research on its etiological characteristics is quite limited