cDNA library Xenopus laevis oocyte

Description

This cDNA library (plasmid DNA) is constructed from Xenopus oocyte-derived poly(A) RNA 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 pBA2 vector used in this library has pUC ori which enables replication in E. coli and Amp^r as a selection marker.

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 level of mRNA of the particular 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: 1.1 x 10⁶

              2) Average insert size : longer than 1 kb

Storage:  -20°C

References

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)

Note

* This library is to be used only by the purchaser.  It is not allowed to amplify and transfer the library to a third person.