cDNA Library S. cerevisiae Log Phase

This cDNA library (plasmid DNA) is constructed from Saccharomyces cerevisiae strain S288C- derived poly(A)+ RNA at the log phase by the Linker-Primer method?(Ref.1) by Prof. H. Nojima of Osaka University. This library is 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 pLZ3 vector (shown below) used in this library can not replicate in S. cereviseaas but contains pUCori for replication in E. coli. 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. 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 rate of mRNA of the objective gene.)