Selective breakage of ONA alongside 5-bromodeoxyuridine nucleotide residues by high temperature hydrolysis
- 1 January 1977
- journal article
- Published by Oxford University Press (OUP) in Nucleic Acids Research
- Vol. 4 (4), 969-987
- https://doi.org/10.1093/nar/4.4.969
Abstract
The substitution of thymine mucleotides (pT) in oligodeoxynucleotides by bromouracil nucleotides (pBU) changes the properties of the oligonucleotides in two ways: (1) It alters their mobility during DEAE-Cellulose homochromatography1. (2) It substantially enhances their sensitivity to high temperature hydrolysis under mildly alkaline conditions (pH 8.9). The resultant breaks occur adjacent to pBU residues and leave terminal phosphates on the breakage products. With more extreme conditions some loss of terminal phosphates can occur. Heating at 100 degrees for 16 hr at pH 8.9 produces cleavage at about half of the pBU residues with minimal loss of terminal phosphates. The properties described here may explain the thermal sensitivity of bacteria grown in 5BU2 and may have a use in DNA sequencing technology.Keywords
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