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Precast Gel protocol

• Notes for using Bio-Rad devices

1. To use Bio-Rad devices, remove the gasket from the inner frame, turn it around so the flat side is facing outwards and re-insert into the inner frame. (Reverse device gasket matching flat side to gel)

how to use Bio Rad devices-01

2. Place the precast gels onto the running frame, and then place the running frame into the tank.

3. Load samples onto gel and run the gel for 1 hour at 140 V in 1x SDS Running Buffer.

• Notes for using Life-Tech devices

A tight seal must be formed between the gel cassette and the gasket of the running frame to prevent leakage. In the NOVEX Tanks, adaptor plates must be used to form a tight seal. (Requires Adaptor Plate to adapt for cassette thickness)

how to use Bio Rad devices-02

1. Two adaptor plates are required when running just one precast gel. Place the precast gel onto the running frame. Place two adaptor plates onto the back of the running frame. (Ensure that the wells are facing away from the inner tank so that buffer does not leak.)

2. Put the running frame into the tank. Using the wedge device, clamp the running frame and the gels into place. The running frame should be fixed tightly and the gels should not be able to move.

3. Load samples onto gel and run the gel for 1 hour at 140v in 1x SDS Running Buffer.

PAGE Gel Solution Protocol

1. Pipet appropriate amount of the resolving gel solution in a small beaker, add freshly made 10% APS at a ratio of 1:25 (v/v). Mix well by vigorously pipetting for 20-30 times or with a magnetic stir bar for 20 seconds. Steadily dispense the resolving gel solution into the cassette.

Proceed to step 2 immediately.

2. Pipet appropriate amount of the stacking gel solution in another small beaker, add freshly made 10% APS at a ratio of 1:25 (v/v). Mix well by vigorously pipetting for 20-30 times or with a magnetic stir bar for 20 seconds. Pour the stacking gel solution on top of the resolving gel (Without waiting for solidification) and insert a gel comb into the stacking gel.

3. Allow the gel to polymerize for 30–50 min. Room temperature (22–25°C) is optimal for polymerization.

4. Load samples onto gel and run the gel for 1 hour at 140v in 1x SDS Running Buffer.

Running voltage change is not recommended. Use Tris-glycine-SDS running buffer or MOPS-SDS running buffer.

• Recommended volume of gel solution (mini gel)

Gel thickness (mm) Resolving gel solution (ml) Stacking gel solution (ml)
0.75 4 1
1.0 5 1.2
1.5 7 2

• Recommended gel percentage (mini gel)

Gel percentage Separation range (kDa) Optimum separation range (kDa)
8% 35-500 36-94
10% 20-350 25-80
12% 10-300 12-60
15% 8-300 10-43
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