Accelerated separation of random complex DNA patterns in gels: comparing the performance of discontinuous and continuous buffers.
Complex DNA fragment patterns produced by random amplified polymorphic DNA (RAPD) assays were separated in agarose as well as polyacrylamide gels using a continuous buffer and two discontinuous buffer systems, with and without urea, respectively. The effect of very high field strength/current levels on the duration of separation, the relative mobilities of the selected bands, and the appearance of the pattern was tested. The use of discontinuous buffer systems resulted in sharper bands and better resolution at 30 min duration of separation for DNA fragments in the size range of 200-2000 base pairs. These observations could help researchers in the rapid separation of band patterns produced by polymerase chain reaction (PCR)-based methods utilized in small- to mid-scale genome searches.
Zsolnai
A
A
Research Institute for Animal Breeding and Nutrition, Herceghalom, Hungary.
Orbán
L
L
D000818:Animals; D001888:Boric Acids / Q000032:analysis; D002021:Buffers; D004247:DNA / Q000032:analysis*; D004492:Edetic Acid / Q000032:analysis; D004553:Electric Conductivity; D004587:Electrophoresis, Agar Gel / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D019105:Random Amplified Polymorphic DNA Technique / Q000295:instrumentation*; D014325:Tromethamine / Q000032:analysis
D001888:Boric Acids; D002021:Buffers; C069591:Tris-borate-EDTA buffer; D014325:Tromethamine; D004247:DNA; D004492:Edetic Acid