Preferential modification of guanine bases in DNA by dimethyldioxirane and its application to DNA sequencing.
From gel sequencing experiments with 32P-end-labelled oligodeoxyribonucleotides, it is shown that treatment of DNA with the powerful oxidant dimethyldioxirane, followed by heating in piperidine, causes selective strand scission at the sites of guanine bases. The same specificity for cleavage at guanine was observed with a 45-mer labelled at either the 3'- or 5'-end and with a single and double stranded 34-mer. On account of its speed and operational simplicity, modification with dimethyldioxirane is proposed as a practicable alternative to conventional chemical sequencing procedures for locating guanine bases in DNA.
Davies
R J
RJ
Division of Biochemistry, School of Biology and Biochemistry, Queen's University, Northern Ireland.
Boyd
D R
DR
Kumar
S
S
Sharma
N D
ND
Stevenson
C
C
D001483:Base Sequence; D004247:DNA / Q000032:analysis* / Q000187:drug effects; D004852:Epoxy Compounds / Q000494:pharmacology*; D004988:Ethers, Cyclic / Q000494:pharmacology*; D006147:Guanine / Q000032:analysis*; D008969:Molecular Sequence Data; D009710:Nucleotide Mapping; D015345:Oligonucleotide Probes
D004852:Epoxy Compounds; D004988:Ethers, Cyclic; D015345:Oligonucleotide Probes; D006147:Guanine; D004247:DNA; C064455:dimethyldioxirane