RNA sequencing using fluorescent-labeled dideoxynucleotides and automated fluorescence detection
Open Access
- 1 January 1990
- journal article
- research article
- Published by Oxford University Press (OUP) in Nucleic Acids Research
- Vol. 18 (4) , 879-884
- https://doi.org/10.1093/nar/18.4.879
Abstract
Although dideoxy terminated sequencing of RNA, using reverse transcriptase and oligodeoxynucleotide primers, is now a well established method, the accuracy is limited by sequence ambiguities due to unspecific chain termination events. A protocol is described which circumvents these ambiguities by using fluorescence labels tagged to dideoxy nucleotides. Only chain terminations caused by dideoxynucleotides were detected while premature terminated cDNA's remain undetectable. In addition, the remaining multiple signals at nucleotide positions can be assigned to sequence heterogeneities within the RNA sequence to be determined.Keywords
This publication has 39 references indexed in Scilit:
- Temporal fluctuations in HIV quasispecies in vivo are not reflected by sequential HIV isolationsCell, 1989
- Use of avian myeloblastosis virus reverse transcriptase at high temperature for sequence analysis of highly structured RNAGene Analysis Techniques, 1989
- An in vivo recombinant RNA capable of autocatalytic synthesis by Qβ replicaseNature, 1988
- CUUCGG hairpins: extraordinarily stable RNA secondary structures associated with various biochemical processes.Proceedings of the National Academy of Sciences, 1988
- A System for Rapid DNA Sequencing with Fluorescent Chain-Terminating DideoxynucleotidesScience, 1987
- A non-radioactive automated method for DNA sequence determinationJournal of Biochemical and Biophysical Methods, 1986
- Resolution of a common RNA sequencing ambiguity by terminal deoxynucleotidyl transferaseAnalytical Biochemistry, 1986
- Rapid determination of 16S ribosomal RNA sequences for phylogenetic analyses.Proceedings of the National Academy of Sciences, 1985
- Secondary structure of the circular form of the Tetrahymena rRNA intervening sequence: a technique for RNA structure analysis using chemical probes and reverse transcriptase.Proceedings of the National Academy of Sciences, 1985
- An extracellular Darwinian experiment with a self-duplicating nucleic acid molecule.Proceedings of the National Academy of Sciences, 1967