cDNA Library S.pombe Meiosis

This cDNA library (plasmid DNA) is constructed from Schizosaccharomyces pombe strain CD16-1(h+/h-) derived poly(A)+ RNA at the state of miosis by the Linker-Primer method (Ref.1) by Professor Hiroshi Nojima of Research Institute for Microbial Diseases Osaka University. cDNAs in this library are unidirectionally cloned by using the oligo (dT)18 linker primer which contains the restriction enzyme site of Not I and BamHI (Bgl II)-Sma I adaptor. The pAP3neo vector used in this library can express the S. pombe genes in mammalian cells as it contains SV40 promoter. It also contains f1 ori which is necessary for ssDNA synthesis and bacteriophage T7 and T3 promoter for RNA synthesis (see Figure). GenBank Accession No. AB003468 Application: PCR screening of known or unknown gene: Prepare the primers for the known or unknown gene (cDNA) and amplify the gene by PCR from this library followed by cloning to an appropriate vector. It is useful for large-scale protein productions and preparation of probes etc. Standard amplifying conditions: 35 cycles of PCR reactions using 10-100 ng of cDNA as a template. (Change the quantity of template and the number of cycles depending on the expression levels of mRNA of the genes.)