Rapid identification of genomic interferon genes by sequencing on phage λ DNA templates

Abstract

A method for rapid screening and identification of a gene or closely related genes from a genomic library by directly sequencing λ phage DNA without laborious intermediate steps and subcloning is presented. The Sanger dideoxy sequencing technique with a synthetic eicosadeoxynucleotide (20-mer) primer and double-stranded DNA template was performed at 50°C rather than a room temperature to avoid the formation of nonspecific bands. Detailed steps of the procedure are described. The technique permitted the rapid and convenient sequencing and identification of multiple human leukocyte interferon genes that had been isolated from a human genomic library in phage λ.