Discontinuous polyacrylamide gradient agarose gels resolve a wide range of restriction fragments and optimize the efficiency of nucleic acid transfer.
A vertical electrophoresis procedure utilizing a discontinuous polyacrylamide gradient agarose gel was developed to resolve DNA fragments ranging in size from over 50 kb to less than 300 bp in length. The gel consisted of a polyacrylamide plug at the base of the gel followed by a gradient of agarose ranging from 0.3 to 0.9%. Restriction fragments migrated shorter distances than in a comparable polyacrylamide-0.3% agarose gel, and small fragments were retained. Southern transfer of DNA fragments from the gradient gel onto nitrocellulose was more efficient than transfer of fragments using a nongradient gel.
Jones
C L
CL
Covalent Technology Corporation, Ann Arbor, Michigan 48106.
Simpson
N J
NJ
Waryas
V L
VL
Pappas
M G
MG
D015152:Blotting, Northern; D015139:Blotting, Southern; D004247:DNA / Q000032:analysis*; D004587:Electrophoresis, Agar Gel; D004591:Electrophoresis, Polyacrylamide Gel; D015183:Restriction Mapping
D004247:DNA