cDNA Library S. cerevisiae Log Phase

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 (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. cerevisiae but contains pUC ori 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.)

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
   Storage: -20°C

References: Construction of this library is described in Supplementary data of Ref.3

1. Kobori M et al ”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. Tougan T, Okuzaki D, Nojima H. Chum-RNA allows preparation of a high-quality cDNA library from a single-cell quantity of mRNA without PCR amplification. Nucleic. Acids Res., 36(15):e92, (2008) PMID:18603591

Note

* This library is to be used only by the purchaser. It is not allowed to amplify and transfer the library to a third person.

* Related products: human tissue specific cDNA libraries and cDNA libraries of model organisms (See HP).

Fig. Structure of pLZ3 and the restriction sites.