cDNA Library, Human contact inhibition state - Cosmo Bio Co.,Ltd.

Antibodies

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cDNA Library, Human contact inhibition state

Catalog No.: BAM-02-727-EX
Size: 500NG
Price: ¥33000
$440
catalog info: Catalog 2012-p211
Storage: -20C
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Immunogen: Human
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: 50 x 106
2) Average insert size : longer than 1 kb

Other:

 Background 
This cDNA library (plasmid DNA) is constructed from poly(A)+ RNA of human TIG-1 cells (fetal lung fibroblasts) at contact inhibition state. It is constru cted by the Li nker-Primer method by Professo r 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 exp ress human genes in m ammalian cells as it cont ains 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 Accession No. AB003468
 Application 
PCR screening of kno wn or u nknown gene: Prepare the primers for the known or unknown gene (cDNA) and amplify the gene by PCR from this library followed by clonin g to an approp riate 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 of the 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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