Conference Proceedings

Mutation detection using fluorescent enzyme mismatch cleavage with T4 endonuclease VII

JJ Babon, M McKenzie, RGH Cotton

ELECTROPHORESIS | WILEY-V C H VERLAG GMBH | Published : 1999

Abstract

Mutation detection techniques are often limited by sensitivity, ease of use and short fragment lengths. Enzyme mismatch cleavage (EMC) is a technique capable of rapidly scanning 1 kbp fragments of DNA for mutations. It relies on the ability of a bacteriophage resolvase enzyme, T4 endonuclease VII, to cleave DNA at single base pair mismatches and small heteroduplex loops. Originally the process was performed using radioactively labeled DNA and the results analysed after denaturing polyacrylamide gel electrophoresis and autoradiography. However, access to systems capable of detecting fluorescent species migrating through a gel and the widespread availability of fluorescently tagged primers hav..

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