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Genomics. 1996 Nov 1;37(3):327-36.

High-efficiency full-length cDNA cloning by biotinylated CAP trapper.

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  • 1Genome Science Laboratory, Tsukuba Life Science Center, Ibaraki, Japan.

Abstract

We have devised a method for efficiently constructing high-content full-length cDNA libraries based on chemical introduction of a biotin group into the diol residue of the cap structure of eukaryotic mRNA, followed by RNase I treatment to select full-length cDNA. The selection occurs by trapping the biotin residue at the cap sites using streptavidin-coated magnetic beads, thus eliminating incompletely synthesized cDNAs. When this method was used to construct a mouse brain full-length cDNA library, our evaluation showed that more than 95% of the total clones were of full length, and recombinant clones could be produced with high efficiency (1.2 x 10(7)/10 micrograms starting mRNA). The analysis of 120 randomly picked clones indicates an unbiased representation of the starting mRNA population.

PMID:
8938445
[PubMed - indexed for MEDLINE]
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