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S.cerevisiae, Log Phase : cDNA Library

Background

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

<reference>
1. Kobori,M., Ikeda,Y., Nara,H., Kato,M., Kumegawa,M., Nojima,H., and Kawashima,H. ” Large scale isolation of osteoclast-specific genes by an improved method involving the preparation of a subtracted cDNA library.” Genes Cells 3: 459-475 (1998) PMID: 9753427
2. Tanaka,S. and Nojima,H. ”Nik1: a Nim1-like protein kinase of S. cerevisiae interacts with the Cdc28 complex and regulates cell cycle progression.” Genes Cells 1, 905-921 (1996) PMID: 9077450
3. Sambrook,J. and Russell,DW. Molecular

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.)

<Specification>
Quantity:500 ng (40 ng/ul, 13ul) in 10 mM Tris-HCl-1mM EDTA (pH 7.5)
Quality:
1) Number of independent clones : 3.6 x 106
2) Average insert size : longer than 1 kb


Fig. Structure of pAP3neo and the restriction sites
Ars is the region required for replication in S. pombe, and Ori is a plasmid origin for replication in E. coli.

Product List

Product Name Cat# Quantity Price

cDNA Library, S.cerevisiae Log Phase

BAM-02-701-EX 500NG

¥ 33,000
$ 440
€ 330

To be used for research only. DO NOT use for human gene therapy or clinical diagnosis.