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3G vector-primer plasmid for constructing full-length-enriched cDNA libraries.

Zheng D, Zhou Y, Zhang Z, Li Z, Liu X

Laboratory of Genetics and Molecular Biology, Northeast Forestry University, Harbin 150040, China.

We designed a 3G vector-primer plasmid for the generation of full-length-enriched complementary DNA (cDNA) libraries. By employing the terminal transferase activity of reverse transcriptase and the modified strand replacement method, this plasmid (assembled with a polydT end and a deoxyguanosine [dG] end) combines priming full-length cDNA strand synthesis and directional cDNA cloning. As a result, the number of steps involved in cDNA library preparation is decreased while simplifying downstream gene manipulation, sequencing, and subcloning. The 3G vector-primer plasmid method yields fully represented plasmid primed libraries that are equivalent to those made by the SMART (switching mechanism at 5' end of RNA transcript) approach.

Published 23 June 2008 in Anal Biochem.
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