PCR amplification of cloned gene from recombinant baculovirus by direct PCR for sequencing

Abstract

The present study describes a sophisticated technique to amplify gene fragment directly from the baculovirus without isolating genome. The study is carried out at FMDRL, IVRI, Bengaluru during January to March 2017. The study was performed with a recombinant baculovirus which was propagated in Trichoplusia ni (Tn5) insect cell lines. Here we PCR amplified the gene fragment directly from the recombinant baculovirus without the need for isolation of genomic DNA and sequenced it. Five out of five PCR fragments sequenced shows no changes in the nucleotide sequence which indicates that there was 100% efficiency of the direct PCR approach for amplifying gene fragment from recombinant baculovirus is high. The use of direct PCR approach for amplifying gene fragment from recombinant baculovirus helps to cut short the time and expenses for amplification of gene fragment from baculovirus for sequencing.