A new PCR based method for the generation of nested deletions.
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.
Whitcomb
J M
JM
ABL-Basic Research Program, NCI-Frederick Cancer Research and Development Center, MD 21702-1201.
Rashtchian
A
A
Hughes
S H
SH
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
D009838:Oligodeoxyribonucleotides; D009843:Oligoribonucleotides; D012313:RNA; D004247:DNA