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A new PCR based method for the generation of nested deletions
J M Whitcomb1, A Rashtchian, S H Hughes
1ABL-Basic Research Program, NCI-Frederick Cancer Research and Development Center, MD 21702-1201.
Abstract:
We have developed a simple, PCR-based protocol, random primed/anchored-PCR (RPA-PCR), that allows the selective amplification and efficient cloning of segments that are adjacent to any known sequence. We demonstrate that RPA-PCR can be used to prepare a nested set of evenly spaced deletions suitable for DNA sequencing. However, it should also be possible to use this technique for a number of other purposes: generating deletions for the analysis of eukaryotic promoters, extending cDNA clones in the 5' direction, cloning the insertion sites of retroviral proviruses and transposons, and analyzing intron/exon boundaries.