Detection of a single base exchange in PCR-amplified DNA fragments using agarose gel electrophoresis containing bisbenzimide-PEG.
Using PCR fragments of known sequences derived from isolates of two related fungal species, simple submarine electrophoresis in agarose gels containing a bisbenzimide-PEG conjugate (H.A.-Yellow) has been shown to be capable of distinguishing DNA fragments 567 bp long which differ by as little as a single base change. However, only changes affecting bisbenzimide binding sites (which consist of at least four consecutive A/T bases) alter mobility; other changes are ineffective. A second ligand (H.A.-Red) with high G/C specificity is suggested which may be as effective in detecting other sequence changes.
Müller
M
M
Hanse Analytik GmbH, Fahrenheitstrasse 1, D-28359 Bremen, Germany. m.mueller@hanse-analytik.de
Kruse
L
L
Tabrett
A M
AM
Barbara
D J
DJ
D006690:Bisbenzimidazole / Q000031:analogs & derivatives*; D004252:DNA Mutational Analysis / Q000379:methods*; D004271:DNA, Fungal / Q000032:analysis*; D004587:Electrophoresis, Agar Gel / Q000379:methods*; D005456:Fluorescent Dyes; D011092:Polyethylene Glycols; D016133:Polymerase Chain Reaction
D004271:DNA, Fungal; D005456:Fluorescent Dyes; C110412:H.A.-yellow; D011092:Polyethylene Glycols; D006690:Bisbenzimidazole