cDNA library Planaria Planaria

This cDNA library (plasmid DNA) is constructed from Planaria-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 (Bgl II)-Sma I adaptor. The pAP3neo vector used in this library can express in mammalian cells as it contains SV40 promoter. It also contains Ori for the plasmid replication and f1 ori which is necessary for ssDNA synthesis in E. coli 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 levelof mRNA of the particular gene.)