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Abstract for Universal RiboClone® cDNA Synthesis System
The Universal RiboClone® cDNA Synthesis System contains the
reagents required for the synthesis of double-stranded cDNA from mRNA, and
subsequent ligation into a suitable vector. The system is based on the method
described by Okayama and Berg, with modifications by Gubler and Hoffman. First
strand synthesis is driven by AMV (Avian Myeloblastosis Virus) Reverse
Transcriptase and either Random Hexameric Primers or an Oligo(dT) Primer,
followed directly by second strand replacement synthesis using RNase H and DNA
Polymerase I. After treatment with T4 DNA Polymerase to flush the ends, the
double-stranded cDNA molecules are prepared for cloning by size fractionation
and the addition of EcoR I Adaptors. The resulting cDNA preparation can
then be cloned into a suitable vector.
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