Random Primed Gene Walking PCR: A Simple Procedure to Retrieve Nucleotide Fragments Adjacent to Known DNA Sequences

摘要

The development of the polymerase chain reaction (PCR) has simplified (he cloning and studying of nucleotide sequences. Modifications of PCR protocols have been used for gene walking. Most of these protocols, such as inverse PCR (5), require multiplesteps. Recently, a few variations of a procedure called targeted gene walking PCR (TGW-PCR) have been described to accomplish the same task (1-3). However, in most cases, TGW-PCR amplifies multiple nonspecific PCR products from which gene-specific PCR products must be distinguished and purified before attempting to sequence them (1-3).