Resolution of DNA molecules greater than 5 megabases by contour-clamped homogeneous electric fields.
Excellent resolution of chromosomal DNA molecules from Saccharomyces cerevisiae, Candida albicans and Schizosaccharomyces pombe has been obtained using alternating contour-clamped homogeneous electric field (CHEF) gel electrophoresis. The largest of these molecules is greater than 5 Mb in size and is resolved after 130 hours in a 0.6% agarose gel at a field strength of 1.3 V/cm and a switching interval of 1 hour. Separation of concatamers of phage lambda DNA reveals four regions of resolution in alternating CHEF gel electrophoresis. There are two regions of good resolution in which mobility approximates a linear function of molecular weight. These are separated by a region of lower resolution and bounded at high molecular weights by a region of little or no resolution. The four regions are of practical and possibly theoretical importance.
D010582:Bacteriophage lambda / Q000032:analysis; D002176:Candida albicans / Q000032:analysis; D002875:Chromosomes / Q000032:analysis; D004247:DNA / Q000032:analysis*; D004271:DNA, Fungal / Q000032:analysis; D004279:DNA, Viral / Q000032:analysis; D004586:Electrophoresis / Q000379:methods*; D004587:Electrophoresis, Agar Gel / Q000379:methods*; D006801:Humans; D008970:Molecular Weight; D012441:Saccharomyces cerevisiae / Q000032:analysis; D012568:Schizosaccharomyces / Q000032:analysis
D004271:DNA, Fungal; D004279:DNA, Viral; D004247:DNA