Construction of normal human lymphocyte cDNA library

Abstract A lymphocyte cDNA library of normal human was constructed in order to obtain specific gene and prepare lymphocyte gene chips to detect the relative genes between psychiatric diseases and immunity. The lymphocyte was abstracted from fresh normal human blood and cultured in vitro. Total RNA of lymphocyte was extracted from the cultured cells and then mRNA was extracted further. Moreover,single-strand cDNA and double-strand cDNA were synthesized in turn. The double-strand cDNAs were ligated to SalI and NotI adaptor,which were later ligated to arms of gammaZipLox. Ligated-cDNAs were packed in vitro, and then infected E. coli Y1090. Titering the phage and amplifying the library. The lymphocyte cDNA library consisted of 2.6 x 10(6) recombinants with the length of 1-5 kb and the cloning efficiency was 5 x 10(12) recombinants/g cDNA. The amplified library was 3 x 10(7) recombinants/microl in concentration and the number of bacteriophage plagues was the most suitable in density after it was diluted to 10(-6) in concentration. The constructed cDNA library of normal human lymphocyte would be helpful to further detecting target genes and preparing gene chips etc.