3G vector-primer plasmid for constructing full-length-enriched cDNA libraries.
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.
Zheng
Dong
D
Laboratory of Genetics and Molecular Biology, Northeast Forestry University, Harbin 150040, China.
Zhou
Yanna
Y
Zhang
Zidong
Z
Li
Zaiyu
Z
Liu
Xuedong
X
D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics*; D015723:Gene Library; D005822:Genetic Vectors / Q000235:genetics*; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics*
D017931:DNA Primers