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 Primers or an Oligo(dT)₁₅ 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 prep...

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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 Primers or an Oligo(dT)₁₅ 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 EcoRI Adaptors. The resulting cDNA preparation can then be cloned into a suitable vector.

Features - Benefits

• Convenient: Contains all the necessary reagents to synthesize double-stranded cDNA from RNA.
• Flexible: Both Oligo(dT)₁₅ Primer and Random Primers are included, providing the researcher a choice of priming methods.

Applications

• Synthesis of double-stranded cDNA from poly(A)+ RNA.

References

1. Okayama, H. and Berg, P. (1982) Mol. Cell Biol. 2, 161–70.
2. Gubler, U. and Hoffman, B.J. (1983) Gene 25, 263–9.

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