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PubMed · 1693515

Optimal conditions for synthesizing complementary DNA in the HIV-1 endogenous reverse transcriptase reaction.

W H Yong·S Wyman·J A Levy
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Abstract

Concentration of monovalent and divalent cations, anionic detergent, reducing agent and nucleotides, as well as pH, temperature, and incubation time were optimized for high levels of HIV-1 endogenous reverse transcriptase activity. In addition, mellitin, a peptide substitute for anionic detergent, and oligo(dT)12-18 were found to stimulate nucleic acid synthesis. This HIV-1 endogenous reaction demonstrated RNA- and DNA-dependent DNA polymerase activities. Nucleic acid intermediates and final products included RNA:DNA hybrids as well as single- and double-stranded DNA. The complementary DNA products formed were representative of all regions of the HIV-1 genome.

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CationsDNAGenes, ViralGenetic TechniquesHIV-1Indicators and ReagentsMelittenNucleic Acid HybridizationOligodeoxyribonucleotidesRNA-Directed DNA PolymeraseTranscription, GeneticJournal ArticleResearch Support, U.S. Gov't, P.H.S.

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W H Yong, S Wyman, J A Levy. 1990. Optimal conditions for synthesizing complementary DNA in the HIV-1 endogenous reverse transcriptase reaction.. https://doi.org/10.1097/00002030-199003000-00004

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