Bio-Rad Trans-Blot® Plus Cell - Manual (Page 17)

Table 3.4 DNA and RNA
These conditions were determined empirically using 5% uniform TBE Criterion
gels and the low range Fluorescein labeled DNA standards. See Section 3.3 for
buffer formulations.
High Intensity Field Conditions (Cooling Required)
PLATE ELECTRODE DISTANCE
4 cm 7 cm 10 cm
Buffer Power Run Power Run Power Run
conditions Time conditions Time conditions Time
1X TBE 30V/2A 30 min 44V/2A 45 min 63V/2A 60 min
40V/3A 15–30 min 67V/3A 15–30 min 93V/3A 15–30 min
1X TAE 21V/2A 30 min 35V/2A 45 min 454V/2A 60 min
30V/3A 15–30 min 50V/3A 15–30 min 70V/3A 15–30 min
Standard Field Conditions (Cooling Recommended)
PLATE ELECTRODE DISTANCE
4 cm 7 cm 10 cm
Buffer Power Run Power Run Power Run
conditions Time conditions Time conditions Time
1X TBE 10V/0.6A 16 Hrs. 10V/0.35A 16 Hrs. 10V/0.25A 16 Hrs.
20V/0.85A 20V/0.55A
30V/0.9A
1X TAE 10V/0.8A 16 Hrs. 10V/0.5A 16 Hrs. 10V/0.35A 16 Hrs.
20V/0.8A
3.2 Advice for Electrophoretic Transfer
1. Equilibration of gels
All electrophoresis gels should be equilibrated in transfer buffer prior to
electrophoretic transfer to remove contaminating electrophoresis buffer salts. If
salts are not removed, they will increase the conductivity of the transfer buffer
and the amount of heat generated during the transfer. Also, gels will shrink or
swell to various degrees in the transfer buffer depending on the acrylamide
percentage and the buffer composition. Equilibration allows the gel to adjust to
its final size prior to electrophoretic transfer. Equilibration is not necessary in
situations where the same buffer is used for both electrophoresis and transfer
(e.g., native gel transfers).
15