Description
This cDNA library (plasmid DNA) is constructed from Xenopus oocyte-derived poly(A)+ RNA by the Linker-Primer method by Professor Hiroshi Nojima of Research Institute for Microbial Diseases, 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 (BglII)-SmaI adaptor. The pBA2 vector used in this library has the pUC ori which enables replication in E. coli and Ampr as a selection marker.
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 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 rate of mRNA of the objective gene.)
