Use of short oligonucleotides to screen cosmid libraries for clones containing G/C-rich sequences |
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Authors: | G Melmer M Buchwald |
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Affiliation: | Research Institute, Hospital for Sick Children, Toronto, Ontario, Canada. |
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Abstract: | We 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 of Not I, was used to screen a cosmid library, and it led to a greater than 10-fold enrichment in the number of clones containing Not I 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. |
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