Rapid Isolation of Full-Length cDNA Sequence from cDNA Library by Hot-Start Polymerase Chain Reaction

Aim To establish a new technique for rapid isolation full length cDNA sequence of human novel gene from cDNA library using hot start PCR. Methods The library vector specific primers and gene specific primers have been designed for performing hot start PCR to attain the full length cDNA of differential expressed sequence tag(FRG4) of U937 foam cell formation induced by ox LDL from human fetal liver cDNA library. The products of PCR are cloned to pGEM T vector and sequenced. Results We have successfully attained the full length cDNA sequence of FRG4. Conclusion This technique is a rapid and efficient method for isolating full length cDNA sequence of human novel gene from cDNA library.