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Research on RT-PCR method for amplifying S1 gene of infectious bronchitis virus isolates

Desheng Chen, Jieyan Pan, Liqun Jia, Dai Ya-bin, Baoxiang Cai, Puyan Chen

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Abstract

According to the Genbank open sequences, a pair of special primers was designed to amplify the whole S1 gene of IBV isolates. M41, the reference strain of IBV, was chosen to optimize the main elements in RT-PCR. The concentrations of Mg~(2+), dNTP and primers were defined to be 1.5 mmol/L, 20 mumol/L and 40 mumol/L respectively after the experimentation. Based on the optimized system, JS.95/03, SD/97/01 and other isolates of IBV in China were amplified. The expected 1.7 kb cDNA fragments of whole S1 gene were obtained. It is conclude that the established method is general to some extent for IBV S1 gene amplifying.

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What this paper is about

According to the Genbank open sequences, a pair of special primers was designed to amplify the whole S1 gene of IBV isolates. M41, the reference strain of IBV, was chosen to optimize the main elements in RT-PCR. The concentrations of Mg~(2+), dNTP and primers were defined to be 1.5 mmol/L, 20 mumol/L and 40 mumol/L respectively after the experimentation. Based on the optimized system, JS.95/03, SD/97/01 and other isolates of IBV in China were amplified. The expected 1.7 kb cDNA fragments of whole S1 gene were obtained. It is conclude that the established method is general to some extent for IBV S1 gene amplifying.

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Available abstract

According to the Genbank open sequences, a pair of special primers was designed to amplify the whole S1 gene of IBV isolates. M41, the reference strain of IBV, was chosen to optimize the main elements in RT-PCR. The concentrations of Mg~(2+), dNTP and primers were defined to be 1.5 mmol/L, 20 mumol/L and 40 mumol/L respectively after the experimentation. Based on the optimized system, JS.95/03, SD/97/01 and other isolates of IBV in China were amplified. The expected 1.7 kb cDNA fragments of whole S1 gene were obtained. It is conclude that the established method is general to some extent for IBV S1 gene amplifying.

Key concepts: Infectious bronchitis virus, GenBank, Biology, Gene, Avian infectious bronchitis virus, Avian infectious bronchitis, Virology, Complementary DNA

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