cDNA Library, S.pombe ; mitosis - Cosmo Bio Co.,Ltd.

Antibodies

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cDNA Library, S.pombe ; mitosis

Catalog No.: BAM-02-703-EX
Size: 500NG
Price: ¥33000
$440
catalog info: Catalog 2012-p211
Storage: -20C
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Immunogen: Schizosaccharomyces pombe
Purity: >98% by SDS-PAGE
Purpose: [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.
component: [Size]
500 ng (40 ng/ ul , 13ul ) in 10 mM Tris-HCl-1mM EDTA (pH 7.5)
[Quality]
1) Number of independent clones: 28 x 106
2) Average insert size: longer than 1 kb

Other:

 Background 
This cDNA library (plasmid DNA) is constructed from Schizosaccharomyces pombe strain h-L972-derived poly(A)+ RNA at the log phase by the Linker-Primer method by Prof. H. Nojima of Research Institute for
Microbial Diseases, Osaka University. cDNAs in this library are unidirectionally cloned by using the oligo (dT)18 linker primer which contains the restriction enzyme sites of Not I, and BamHI (Bgl II)-Sma I adaptor.
The pLZ3 vector used in this library can replicate both in S. pombe and E.coli, and express the S. pombe genes in mammalian cells as it contains SV40 promoter as well as in S. pombe.
 Application 
1. PCR screening of known or unknown gene: Prepare the primers for the known or unknown gene (cDNA) and amplify thegene by PCR from this library followed by cloning to an appropriate vector. The cloned cDNAs are useful for identifying the coding region, large-scale protein production, 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 levels of mRNA of the gene of interest.)
2. Cloning the cDNA by functional complementation of the correspondingS. pombe mutants.



 IMAGE1



Fig. Structure of pLZ3 and the restriction sites Ars is the S. pombe region required for replication in S. pombe, and Ori is a plasmid origin for replication in E. coli

Reference:

•  Kobori M et al., Genes Cells 3: 459-475 (1998)
•  Tanaka S. et al., Genes Cells 1, 905-921 (1996)
•  Sambrook J. et al., CSHL Press (2001)
 
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