cDNA Library, Rat NRK Cell Log Phase - Cosmo Bio Co.,Ltd.

Antibodies

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cDNA Library, Rat NRK Cell Log Phase

Catalog No.: BAM-02-719-EX
Size: 500NG
Price: ¥33000
$440
catalog info: Catalog 2012-p211
Storage: -20C
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Immunogen: Rat
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: 1.4 x 106
2) Average insert size : longer than 1 kb

Other:

 Background 
This cDNA library (plasmid DNA) is constructed from poly(A)+ RNA of rat NRK (normal rat kidney) cells at the 0hr log phase. It is constructed by the Linker-Primer method by Professor Hiroshi Nojima of Research
Institute for Microbial Diseases, 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 pAP3neo vector used in this library can express ratgenes in mammalian cells as it contains SV40 promoter. It also contains Ori of pUC plasmid required for replication in E.coli, f1 ori which is necessary for ssDNA synthesis, and bacteriophage T7 and T3 promoters for RNA synthesis. GenBank AccessionNo. 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 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 rate of mRNA ofthe objective gene.)



 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)
 
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