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 th e utilization of native polyacrylamide gels (without urea) could speed up t he separation of polymerase chain reaction (PCR)-amplified products with an automated 377 DNA sequencer, denatured PCR products were subjected to elec trophoresis in 6% native gels under 45 degreesC. Results show that a decrea se of the electrophoresis temperature from 51 degreesC (recommended by the User's Manual) to 45 degreesC substantially facilitates the preservation of gel wells, and that all PCR products tested migrate significantly faster i n native than in denatured (with urea) gels of the same concentration. The combination of a 6% native gel and a lower (45 degreesC) electrophoresis te mperature permits multiple uses of a given gel with consistent results, con sequently reducing the electrophoresis time and reagent costs.