[New method for the synthesis and cloning of cDNA].
A simple method for cloning cDNA has been suggested. The plasmid pUC18 was digested with Pst1. A plasmid primer for cDNA synthesis was prepared by dT tailing with terminal transferase. After synthesis of cDNA, dG tails were added and then 3' ends blocked with rG. The plasmid was digested with Kpn1 and dC tails were added, after which annealing took place and RNA:DNA hybrids were used for Escherichia coli transformation. The efficiency of approx. 10(4) transformants per microgram of starting mRNA has been obtained.
D003001:Cloning, Molecular / Q000187:drug effects / Q000379:methods*; D004247:DNA / Q000096:biosynthesis*; D004262:DNA Restriction Enzymes / Q000494:pharmacology; D004274:DNA, Recombinant; D004926:Escherichia coli / Q000235:genetics; D009693:Nucleic Acid Hybridization / Q000187:drug effects; D010957:Plasmids / Q000187:drug effects; D012333:RNA, Messenger / Q000235:genetics; D014169:Transformation, Bacterial
D004274:DNA, Recombinant; D012333:RNA, Messenger; D004247:DNA; D004262:DNA Restriction Enzymes
Novyĭ metod sinteza i klonirovaniia kDNK.