Description
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 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 NotI and BamHI (BglII)-SmaI adaptor. The pLZ3 vector used in this library cannot replicate in S. cerevisiae but contains the pUC ori for replication in E. coli.
Application 1: PCR cloning of known or unknown genes.
Prepare primers for known or unknown genes (cDNAs) and amplify by PCR from this library followed by cloning into 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 mRNA expression levels of the gene of interest.)
