Identifying tagged transposon insertion sites in yeast by direct genomic sequencing.
Tagged transposons are powerful tools for large-scale studies of gene expression, protein localization, and gene disruption in Saccharomyces cerevisiae. The current techniques used to identify transposon insertion sites in the yeast genome require a DNA amplification step that can be time-consuming and problematic. We show that the DNA amplification step can be bypassed. Insertion sites can be identified rapidly and reliably by direct genomic sequencing using a transposon-specific primer, BigDye-labelled terminators, and an automated sequencer. Direct genomic sequencing can also save time on the genetic analysis phase of transposon-based projects.
Horecka
J
J
Department of Molecular Biology, NIBH, AIST, 1-1 Higashi, Tsukuba, Ibaraki 305-8566, Japan. horeka@nibh.go.jp
Jigami
Y
Y
D001483:Base Sequence; D004251:DNA Transposable Elements; D016681:Genome, Fungal; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D017422:Sequence Analysis, DNA
D004251:DNA Transposable Elements