title: A new PCR based method for the generation of nested deletions.
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.
authors.affiliation: ABL-Basic Research Program, NCI-Frederick Cancer Research and Development Center, MD 21702-1201.
authors.forename: J M
authors.initials: JM
authors.lastname: Whitcomb
authors.forename: A
authors.initials: A
authors.lastname: Rashtchian
authors.forename: S H
authors.initials: SH
authors.lastname: Hughes
mesh_terms: D001483:Base Sequence; D003001:Cloning, Molecular; D004247:DNA; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D009843:Oligoribonucleotides; D016133:Polymerase Chain Reaction / Q000379:methods*; D012313:RNA; D017384:Sequence Deletion
chemical_list: D009838:Oligodeoxyribonucleotides; D009843:Oligoribonucleotides; D012313:RNA; D004247:DNA