cDNA library S.pombe Mitosis

This cDNA library (plasmid DNA) is constructed from Schizosaccharomyces pombe strain h-L972- derived poly(A)+ RNA at the log phase by the Linker-Primer method (Ref.1) by Prof. H. 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 sites of Not I and BamHI (Bgl II)-Sma I adaptor. The pLZ3 vector used in this library can replicate both in S. pombe and E.coli and express the S. pombe genes in mammalian cells as it contains SV40 promoter as well as in S. pombe. 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. The cloned cDNAs are useful for identifying the coding region large-scale protein production 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 gene of interest.)