Determination of DNA base composition by small scale acrylamide-CsCl gradient centrifugation.
In this paper we describe a simple and rapid protocol for DNA base composition determination by CsCl gradient in the presence of acrylamide. This method permits the determination of GC content in microgram amounts of DNA, and results are easily documented in photographs or graphs. The protocol was applied to the characterization of nematode DNA, but can be used for other organisms. Analyzing several experiments the mean standard deviation observed in the calculated GC content is near 1.3%.
Ahn
Il-Young
IY
Department of Parasitology, Instituto de Ciências Biomédicas, Universidade de São Paulo, Av. Prof. Lineu Prestes, 1374, 05508-900 São Paulo, Brazil.
Winter
Carlos E
CE
D020106:Acrylamide / Q000494:pharmacology; D000818:Animals; D001482:Base Composition; D001703:Biophysics / Q000379:methods*; D017173:Caenorhabditis elegans; D002499:Centrifugation, Density Gradient / Q000379:methods*; D002586:Cesium / Q000494:pharmacology*; D002712:Chlorides / Q000494:pharmacology*; D004247:DNA / Q000737:chemistry / Q000378:metabolism; D004926:Escherichia coli / Q000378:metabolism; D006358:Hot Temperature; D013696:Temperature; D014461:Ultracentrifugation
D002712:Chlorides; D002586:Cesium; D020106:Acrylamide; D004247:DNA; C028019:cesium chloride