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Highly selective chemical modification of cruciform loops by diethyl pyrocarbonate.

Nucleic acids research (1986-05-27)
J C Furlong, D M Lilley
ABSTRACT

Diethyl pyrocarbonate reacts with the single-stranded loops of cruciform structures with great selectivity. Adenine bases are carbethoxylated, as a result of which the backbone may be cleaved with piperidine, and the level of chemical modification at each base may be determined. We have studied the ColE1 and (A-T)34 cruciforms of pColIR315 and pXG540. In each case we observe maximal modification at the most central adenosine of the loop, and an overall pattern of modification corresponding to a total loop size of about six bases. The results may be interpreted in terms of a model in which the loop has a defined tertiary structure. No modification was detected at either cruciform four-way junction, suggesting that this region is fully base-paired.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Diethyl pyrocarbonate, 96% (NT)