Novel display of knotted DNA molecules by two-dimensional gel electrophoresis.
We describe a two-dimensional agarose gel electrophoresis procedure that improves the resolution of knotted DNA molecules. The first gel dimension is run at low voltage, and DNA knots migrate according to their compactness. The second gel dimension is run at high voltage, and DNA knots migrate according to other physical parameters such as shape and flexibility. In comparison with one-dimensional gel electrophoresis, this procedure segregates the knotted DNA molecules from other unknotted forms of DNA, and partially resolves populations of knots that have the same number of crossings. The two-dimensional display may allow quantitative and qualitative characterization of different types of DNA knots simply by gel velocity.
Trigueros
S
S
Institut de Biologia Molecular de Barcelona, CSIC, Jordi Girona 18-26, 08034 Barcelona, Spain.
Arsuaga
J
J
Vazquez
M E
ME
Sumners
D W
DW
Roca
J
J
D003090:Coliphages / Q000235:genetics*; D004250:DNA Topoisomerases, Type II / Q000378:metabolism; D004279:DNA, Viral / Q000737:chemistry* / Q000302:isolation & purification* / Q000378:metabolism; D004587:Electrophoresis, Agar Gel / Q000379:methods*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D009690:Nucleic Acid Conformation; D018583:Pliability; D012680:Sensitivity and Specificity; D015003:Yeasts / Q000201:enzymology
D004279:DNA, Viral; D004250:DNA Topoisomerases, Type II