Multiple use of slab gels in sequencing apparatus for separation of polymerase chain reaction products.
Attempting to assess whether a decrease of the electrophoresis temperature could prevent or reduce the extent of gel well deformations, and whether the utilization of native polyacrylamide gels (without urea) could speed up the separation of polymerase chain reaction (PCR)-amplified products with an automated 377 DNA sequencer, denatured PCR products were subjected to electrophoresis in 6% native gels under 45 degrees C. Results show that a decrease of the electrophoresis temperature from 51 degrees C (recommended by the User's Manual) to 45 degrees C substantially facilitates the preservation of gel wells, and that all PCR products tested migrate significantly faster in native than in denatured (with urea) gels of the same concentration. The combination of a 6% native gel and a lower (45 degrees C) electrophoresis temperature permits multiple uses of a given gel with consistent results, consequently reducing the electrophoresis time and reagent costs.
Man
Y G
YG
Department of Gynecologic and Breast Pathology, Armed Forces Institute of Pathology, American Registry of Pathology, Washington, DC 20306-6000, USA. man@afip.osd.mil
Kuhls
E A
EA
Bratthauer
G L
GL
Moinfar
F
F
Tavassoli
F A
FA
D004247:DNA / Q000737:chemistry / Q000302:isolation & purification; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D005782:Gels; D016133:Polymerase Chain Reaction / Q000379:methods*; D017422:Sequence Analysis, DNA / Q000379:methods*; D013997:Time Factors
D005782:Gels; D004247:DNA