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

Antibodies

>

cDNA Library, S.pombe ; Meiosis

Catalog No.: BAM-02-705-EX
Size: 500NG
Price: ¥33000
$440
catalog info: Catalog 2012-p211
Storage: -20C
@
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: 1.3 x 106
2) Average insert size: longer than 1 kb

Other:

 Background 
This cDNA library (plasmid DNA) is constructed from Schizosaccharomyces pombe, strain CD16-1(h+/h-) derived poly(A)+ RNA at the state of miosis by the Linker-Primer method by Professor Hiroshi Nojima of Research Institute for MicrobialDiseases, Osaka University. cDNAs in this library are 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 canexpress the S. pombe genes in mammalian cells as it contains SV40 promoter. It also contains f1 ori which is necessary for ssDNA synthesis, and bacteriophage T7 and T3
promoter for RNA synthesis. 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 ofprobes, 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 genes.)



 IMAGE1



Fig. Structure of pAP3neo and the restriction sites

Reference:

•  Kobori M et al., Genes Cells 3: 459-475 (1998)
•  Sambrook J. et al., CSHL Press (2001)
 
>