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Direct chemical method for sequencing RNA.
Open accessAbstract
Four different base-specific chemical reactions generate a means of directly sequencing RNA terminally labeled with 32P. After a partial, specific modification of each kind of RNA base, an amine-catalyzed strand scission generates labeled fragments whose lengths determine the position of each nucleotide in the sequence. Dimethyl sulfate modifies guanosine. Diethyl pyrocarbonate attacks primarily adenosine. Hydrazine attacks uridine and cytidine, but salt suppresses the reaction with uridine. In all cases, aniline induces a subsequent strand scission. The electrophoretic fractionation of the labeled fragments on a polyacrylamide gel, followed by autoradiography, determines the RNA sequence. RNA labeled at the 3' end yields clean cleavage patterns for each purine and pyrimidine and allows a determination of the entire RNA sequence out to 100-200 bases from the labeled terminus.
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Citations
Cited by
Oxidative Nucleobase Modifications Leading to Strand Scission
Burrows, C. J., & Muller, J. G. (1998) · Chemical Reviews · doi:10.1021/cr960421s · Cited by 1,736
Interaction of antibiotics with functional sites in 16S ribosomal RNA
Moazed, D., & Noller, H. F. (1987) · Nature · doi:10.1038/327389a0 · Cited by 1,150
METTL1 Promotes let-7 MicroRNA Processing via m7G Methylation
Pandolfini, L., Barbieri, I., Bannister, A. J., Hendrick, A., Andrews, B., Webster, N., Murat, P., Mach, P., Brandi, R., Robson, S. C., et al. (2019) · Molecular Cell · doi:10.1016/j.molcel.2019.03.040 · Cited by 491
“This reaction is the basis of direct RNA sequencing (RNA-seq) by the Maxam and Gilbert method (Peattie, 1979) and has been used formappingm7G residues in highly abundant rRNAs and tRNAs at single nucleotide resolution (Zueva et al., 1985).”
MethodSequence requirements of the hammerhead RNA self-cleavage reaction
Ruffner, D. E., Stormo, G. D., & Uhlenbeck, O. C. (1990) · Biochemistry · doi:10.1021/bi00499a018 · Cited by 470
Pitsch, S., Wendeborn, S., Jaun, B., & Eschenmoser, A. (1993) · Helvetica Chimica Acta · doi:10.1002/hlca.19930760602 · Cited by 137
Metadata
- Authors
- Peattie, D. A.
- Venue
- Proceedings of the National Academy of Sciences
- Published
- 1979-04
- Volume
- 76
- Issue
- 4
- Pages
- 1760-1764
- ISSN
- 0027-8424 (ISSN-L)
- 1091-6490
- OpenAlex
- W2053212888
- Cited by
- 940
- References
- 0
- FWCI
- 30.98
- Type
- journal article
- Language
- en
- Open access
- green
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