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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

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.)

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℃

1. KoboriM. IkedaY. NaraH. KatoM. KumegawaM. NojimaH. and KawashimaH. ” 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. TanakaS. and NojimaH. ”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. SambrookJ. and RussellDW. Molecular Cloning Chapter 11 ”Preparation of cDNA libraries and gene identification.” CSHL Press (2001)
500 ng
330 USD

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