title: RT-PCR method for selective detection of silent gene transcripts in silencing mutants in homothallic strains of Schizosaccharomyces pombe.
abstract: Here we describe a method that allows selective detection of silent copy transcripts in homothallic strains of Schizosaccharomyces pombe in the presence of the active cassettes. The method involving RT-PCR (reverse transcriptase polymerase chain reaction) exploits our observation that the silent copy transcripts extend beyond the regions of homology to the flanking sequences specific for the donor cassettes, thus allowing design of oligos that are specific for the different donors. The results are validated using a known silencing mutant swi6.
authors.affiliation: Institute of Microbial Technology, Chandigarh, India.
authors.forename: Shakil
authors.initials: S
authors.lastname: Ahmed
authors.forename: Jagmohan
authors.initials: J
authors.lastname: Singh
mesh_terms: D002868:Chromosomal Proteins, Non-Histone / Q000235:genetics; D015966:Gene Expression Regulation, Fungal / Q000235:genetics*; D020868:Gene Silencing; D049770:Genes, Mating Type, Fungal / Q000235:genetics; D009154:Mutation / Q000235:genetics*; D012331:RNA, Fungal / Q000032:analysis* / Q000235:genetics; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012568:Schizosaccharomyces / Q000235:genetics*; D029702:Schizosaccharomyces pombe Proteins / Q000235:genetics
chemical_list: D002868:Chromosomal Proteins, Non-Histone; D012331:RNA, Fungal; D012333:RNA, Messenger; D029702:Schizosaccharomyces pombe Proteins; C465958:Swi6 protein, S pombe