In this study, electrophoresis was performed on an ABI Prism 377 (96 well) instrument, using 36, 64, and 96 well gels. Both, 36 and 48-cm well-to-read gels were used. For Dye Primer Chemistry, a 4.25% polyacrylamide 19:1 (6M urea) formulation was used (Anachem, England), whereas for BigDye Terminator chemistry a 5% 29:1 polyacrylamide (6M urea) formulation (Amresco, England) was used. The degased PA formulation was aliquoted and stored at -20℃C.
For 50ml of gel mixture, 250 µl freshly made 10% APS and 35 µl TEMED were added. Gel plates were thoroughly cleaned and mounted in the gel pouring cassette. After casting the gels (using approx. 30 ml of gel mixture for 36-cm gels and 35 ml for 48-cm gels) they were allowed to polymerise for a minimum of 2 hours at room temperature. Gels were always used on the same day.
© 2001 Alexander Binder