Adapted the Sanger chain-termination method for RNA sequencing using reverse transcriptase, successfully sequencing 166 residues of the EMC virus RNA.
Validates RT-based Sanger sequencing in animal virus models; leaves open extension to clinical virology.
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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Zimmern et al. (1978) studied this question.
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