An efficient DNA sequencing strategy based on the bacteriophage mu in vitro DNA transposition reaction.

ABSTRACT: A highly efficient DNA sequencing strategy was developed on the basis of the bacteriophage Mu in vitro DNA transposition reaction. In the reaction, an artificial transposon with a chloramphenicol acetyltransferase (cat) gene as a selectable marker integrated into the target plasmid DNA containing a 10.3-kb mouse genomic insert to be sequenced. Bacterial clones carrying plasmids with the transposon insertions in different positions were produced by transforming transposition reaction products into Escherichia coli cells that were then selected on appropriate selection plates. Plasmids from individual clones were isolated and used as templates for DNA sequencing, each with two primers specific for the transposon sequence but reading the sequence into opposite directions, thus creating a mini

SUBMITTER: Haapa S 

PROVIDER: S-EPMC310728 | biostudies-literature | 1999 Mar

REPOSITORIES: biostudies-literature