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. 1979 Sep 25;7(2):453–464. doi: 10.1093/nar/7.2.453

The synthesis of oligodeoxyribonucleotides using RNA ligase.

D M Hinton, R I Gumport
PMCID: PMC328028  PMID: 386284

Abstract

T4 RNA ligase catalyzes the addition of a single deoxyribonucleoside 3',5'-bisphosphate to the 3'-hydroxyl of oligodeoxyribonucleotides (Hinton et al. (1978) Biochemistry 17, 5091). We have determined improved conditions for this reaction which give yields equal to or greater than 85% when any of five common deoxyribonucleoside bisphosphate (pdAp, pdCp, pdGp, pdTp, or pdUp) are added to dA(PDA)4. A low ATP concentration, which is constantly maintained by a regeneration system composed of phosphocreatine, creatine kinase, and myokinase, contributes to the attainment of high yields. The addition of RNase A and spermine also enhances the rates and yields of the reactions. These conditions facilitate the use of RNA ligase as a reagent for the stepwise synthesis of DNA of defined sequence.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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