Action
Step
Layer 2: Place the two pre-wetted blotting papers on the sponge
and press gently to remove any air bubbles.
Note:
Use a pipette or stirring rod to gently roll out any air bubbles
trapped while assembling the stack.
6
Layer 3: Place the pre-wetted membrane on layer 2 and remove
all air-bubbles.
7
Equilibrate the gel in cold transfer buffer for 10 to 20 minutes.
8
Layer 4: Place the gel on the membrane and ensure there are
no air bubbles.
9
Layer 5: Cover the gel with two pre-wetted sheets of blotting
paper.
10
Layer 6: Finally place a sponge of 3 mm and again press gently
to remove any air bubbles.
11
Close the cassette and press lightly to lock the tabs.
12
Place the cassette into the tank.
13
Run the transfer according to the recommendations from the
transfer tank supplier.
14
Recommended running conditions for transfer
Value
Parameter
25 V
(maximum 400 mA)
Voltage
4°C
Temperature
2.5 hours (with constant stirring)
Time
Amersham ECL Gel and Gel Box User Manual 28-9943-33 AC
55
5 Protocols
5.4 Transfer protocol
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