Use of Short Oligonucleotides to Screen Cosmid Libraries for Clones Containing G/C-Rich Sequences

摘要

ABSTRACTWe have developed a method to identify clones containing recognition sequences for enzymes that cut mammalian genomes infrequently by direct screening of genomic libraries. The degenerate oligonucleotide NNGCGGCCGCNN, in which the internal 8 bases correspond to the recognition sequence ofNotI, was used to screen a cosmid library, and it led to a greater than 10-fold enrichment in the number of clones containingNotI sites. This technique permits the efficient identification of sufficient clones from a chromosome-specific library to allow the construction of a complete pulsed-field map of that chromosome and to assist in finding genes in genomic DNA.