Improved separation of multi-locus hypervariable DNA restriction fragments by field inversion gel electrophoresis and fragment detection using a biotinylated probe.
DNA probes to multiple hypervariable human genomic loci are potentially highly informative as regards individual genome identification and parentage studies. However the resultant Southern blot patterns are generally highly complex and their interpretation difficult, with one important limitation being the extent of fragment separation and resolution. The separation of large DNA restriction fragments in agarose gels may be improved, in comparison to separations obtained by conventional electrophoresis, by the use of field inversion techniques. This is demonstrated by the analysis of the complex human Satellite III related DNA polymorphism. Detection of the Satellite III related restriction fragments is achieved either by using a [35S]-labelled probe (228S) or by using the same probe in a convenient non-isotopic form constructed by the photobiotin process. In addition, the probe 228S is useful for sexing the human genome, by the identification of a Y-specific restriction fragment.
Craig
J
J
School of Biological Sciences, Flinders University of South Australia, Bedford Park.
Fowler
S
S
Skinner
J D
JD
Burgoyne
L A
LA
McInnes
J L
JL
D001710:Biotin; D015139:Blotting, Southern; D004247:DNA / Q000032:analysis*; D015342:DNA Probes; D004587:Electrophoresis, Agar Gel / Q000379:methods; D006801:Humans; D009693:Nucleic Acid Hybridization; D012150:Polymorphism, Restriction Fragment Length
D015342:DNA Probes; D001710:Biotin; D004247:DNA