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Nucleic Acids Research, 1978, Vol. 5, No. 2 615-622
© 1978

Articles

Base-specific reactions useful for DNA sequencing: methylene blue – sensitized photooxidation of guanine and osmium tetraoxide modification of thymine

Theodore Friedmann+ and Daniel M. Brown++

+MRC Laboratory of Molecular Biology, Cambridge, UK and Department of Pediatrics, School of Medicine, University of California at San Diego La Jolla, CA 92093, USA ++University Chemical Laboratory Cambridge, UK

Received January 13, 1978. Exposure of DNA to methylene blue and visible or ultraviolet light causes guanine-specific modification, and subsequent treatment with piperidine leads to chain cleavage at each guanine residue. Treatment of DNA with osmium tetraoxide in dilute pyridine leads to thymidine-specific modification, and subsequent treatment with piperidine leads to chain cleavage at the modified thymidine residues. Both reactions can be used in conjunction with other base specific modifications described by Maxam and Gilbert (1) for the determination of the nucleotide sequence in DNA.


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