Piggybac transposon-based insertional mutagenesis for the fission yeast Schizosaccharomyces pombe

Abstract

Transposon-mediated insertional mutagenesis is a powerful tool for genetic screens. We have developed a piggyBac transposon-based mutagenesis system for the fi ssion yeast Schizosaccharomyces pombe . Here, we describe in detail the procedure for inducing and selecting transpositions, and two protocols for identifying the transposon insertion sites, one using inverse PCR to identify the insertion sites in individual mutants, and the other using high-throughput sequencing to reveal the insertion sites in a mutant pool containing hundreds to thousands of mutants.