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Bio-Rad Trans-Blot® Plus Cell User Manual

Page 17

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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).

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