FAST PULSED-FIELD MINIGEL ELECTROPHORESIS OF LARGE DNA-MOLECULES

Pulsed field gel electrophoresis runs were performed in small Contour Clamped Homogeneous Electric Field (miniCHEF) and Transverse Alternati ng Electric Field (miniTAFE) chambers. Minichambers use minigels and t hey could be suitable for analytical and preparative purposes. In mini chambers higher electric fields and equal heat evolution were achieved with the same voltage applied to conventional chambers. Nine to ten S accharomyces cerevisiae chromosomes were resolved in minichambers appl ying 13-14 V/cm and 50 s of pulse time during 3.2 - 5 h. It was found a linear relationship between the migration per pulse and the electric field (at constant pulse time). An equation relating the pulse time a nd the electric fields was attained. It was used to obtain isopatterns at several electric fields despite the chamber dimensions. Miniappara ti use simple power supplies and cooling systems and save samples, rea gents, laboratory bench space and time.