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Rat NRK Cell, Log Phase : cDNA Library

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 (Ref.1) 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 rat genes 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 (see Figure).

GenBank Accession No. AB003468

<reference>
1. Kobori M et al ” 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. Sambrook J and Russell DW Molecular Cloning Chapter 11 ”Preparation of cDNA libraries and gene identification.” CSHL Press (2001)

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 of the objective gene.)

<Specification>
Quantity :  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


Fig. Structure of pAP3neo and the restriction sites.

Product List

Product Name Cat# Quantity Price

cDNA Library, Rat NRK Cell Log Phase

BAM-02-719-EX 500NG

¥ 33,000
$ 440
€ 330

cDNA Library, S.cerevisiae Log Phase

BAM-02-701-EX 500NG

¥ 33,000
$ 440
€ 330

cDNA Library, S.pombe ; mitosis

BAM-02-703-EX 500NG

¥ 33,000
$ 440
€ 330

cDNA Library, S.pombe ; Meiosis

BAM-02-705-EX 500NG

¥ 33,000
$ 440
€ 330

cDNA Library, S.pombe ; After + HU, (GAMMA), MMS h-L972

BAM-02-707-EX 500NG

¥ 33,000
$ 440
€ 330

cDNA Library, Planaria ; Planaria

BAM-02-709-EX 500NG

¥ 33,000
$ 440
€ 330

cDNA Library, Xenopus ; oocyte

BAM-02-711-EX 500NG

¥ 33,000
$ 440
€ 330

cDNA Library, Mouse ; Thymocyte

BAM-02-713-EX 500NG

¥ 33,000
$ 440
€ 330

cDNA Library, Mouse ; Testis

BAM-02-715-EX 500NG

¥ 33,000
$ 440
€ 330

cDNA Library, Rat ; Rat embryonic fibroblast

BAM-02-717-EX 500NG

¥ 33,000
$ 440
€ 330

cDNA Library, Human ; Umbilical Vein Endothelial Cells

BAM-02-721-EX 500NG

¥ 33,000
$ 440
€ 330

cDNA Library, Human ; HeLa (#1)

BAM-02-723-EX 500NG

¥ 33,000
$ 440
€ 330

cDNA Library, Human ; Fibroblast Primary culture

BAM-02-725-EX 500NG

¥ 33,000
$ 440
€ 330

cDNA Library, Human contact inhibition state

BAM-02-727-EX 500NG

¥ 33,000
$ 440
€ 330

cDNA Library, Human ; 293T

BAM-02-729-EX 500NG

¥ 33,000
$ 440
€ 330

To be used for research only. DO NOT use for human gene therapy or clinical diagnosis.

< Reagents for Genetic Engineerring >