S. Mouradian et al., SEPARATING FIELD-STRENGTH, TEMPERATURE, AND PULSING EFFECTS IN PULSED-FIELD ELECTROPHORESIS, Electrophoresis, 15(8-9), 1994, pp. 1084-1090
The utility of pulsed field electrophoresis for DNA sequencing is inve
stigated. Previous studies have indicated a beneficial retardation of
sequencing fragments when pulsed fields are employed. The interpretati
on of these results is complicated, however, by concomitant variations
in electric field strength and/or temperature. Methods are presented
here permitting discrimination of such mobility effects due to pulsing
, field strength, and temperature. It is demonstrated that under the c
onditions employed here, observed mobility effects are due to electric
field variations rather than pulsing. These conditions thus correspon
d to the low frequency/small molecule limit. The effect of temperature
is estimated from the steady state solution to the heat conduction eq
uation under appropriate boundary conditions. No temperature effect up
on mobility is operative in the thin gel system employed, due to the h
igh efficiency of heat transfer. However, it is shown that in conventi
onal gel systems large temperature-related mobility effects can occur.
These methods for dissecting and understanding mobility effects in pu
lsed field electrophoresis are expected to be of general utility.