September 1, 1978Open Access

3'-terminal nucleotide sequence of encephalomyocarditis virus RNA determined by reverse transcriptase and chain-terminating inhibitors.

Structured PICO

Population

Genomic RNA of the picornavirus encephalomyocarditis virus (EMC virus) and poliovirus RNA

Intervention

Chain-termination method using reverse transcriptase and chain-terminating inhibitors (2',3'-dideoxynucleoside triphosphates) with primer pdT7rC

Outcome

Nucleotide sequence of the 3'-terminal region

Adapted the Sanger chain-termination method for RNA sequencing using reverse transcriptase, successfully sequencing 166 residues of the EMC virus RNA.

Abstract

We have adapted the chain-termination method for determining the nucleotide sequence of DNA of Sanger, Nicklen, and Coulson (1977) Proc. Natl. Acad. Sci. USA 74, 5463--5467 for use with reverse transcriptase (RNA-directed DNA nucleotidyltransferase) on RNA templates. With this method and using a primer (the octanucleotide pdT7rC) directed at the 3'-terminal poly(A), we have determined a sequence of 166 residues in the genomic RNA of the picornavirus encephalomyocarditis virus.

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