On-line melting of double-stranded DNA for analysis of single-stranded DNA using capillary electrophoresis.
Capillary electrophoresis (CE) is a convenient, fast and non-radioactive method with possibilities for automatization. To analyse single-stranded DNA molecules in a more automated way, we developed a heating device to melt double-stranded DNA fragments in the capillary during electrophoresis. In this study we used this device to obtain single-stranded DNA, necessary for the detection of point mutations in DNA using the single-strand conformation polymorphism technique. Results show that double-stranded DNA molecules can be melted on-line into single-stranded DNA molecules, although not for 100%. In an attempt to find universal electrophoretic conditions for the analysis of single-stranded DNA, we investigated the influence of several parameters on the yield of single-stranded DNA molecules and on the resolution of the single-stranded DNA peaks. We demonstrate that this heating device is a technical adjustment of CE which contributes to more automated analyses of DNA fragments.
Kuypers
A W
AW
Department of Hematology, University Hospital Nijmegen, Netherlands.
Linssen
P C
PC
Willems
P M
PM
Mensink
E J
EJ
D004247:DNA / Q000737:chemistry* / Q000235:genetics; D004277:DNA, Single-Stranded / Q000032:analysis* / Q000235:genetics; D019075:Electrophoresis, Capillary / Q000379:methods*; D016158:Genes, p53; D011905:Genes, ras; D017354:Point Mutation; D016133:Polymerase Chain Reaction
D004277:DNA, Single-Stranded; D004247:DNA