Study on Full Length cDNA Library Construction

A simple method of full length cDNA library construction based on long-distance PCR was established.Total RNA from the human placenta tissues were isolated,and the first cDNA was synthesized through RT-PCR with the help of M-MLV reverse enzyme.Then double strand cDNA(ds-cDNA) with the restriction sites of SfiⅠ was synthesized through LD-PCR(Long-distance-PCR) and ligated into the JG45 vector with the restriction sites of Sfi I.Recombinant vectors were transformed into E.coli JM109 and then amplified.Then qualities of the cDNA library were analyzed.The average capacities of the cDNA library was about 7.01×105 clones per μg ds-cDNA with recombinant rate of 96%.Among the 80 detected randomly selected cDNA clones,no repetitive sequence was found.cDNA library constructed with this method was suitable for functional gene analysis.