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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.