2 loading samples, 3 comb volume/maximum well volume, 4 electrophoresis – C.B.S. Scientific C-DASG-400-50 User Manual

Page 11

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Mega-Gel Instructions 12/11

9. Connect the DC power leads to the power supply with the proper polarity. Make sure the black leads are connected

to the black cathode (-) and the red leads are connected to the red anode (+).

10. Rinse sample wells with upper reservoir buffer. Before loading samples, pre-electrophorese the gel for 20 minutes.

WARNING:

Excessive power will cause the gel to overheat and crack the glass plates.

4.2 Loading Samples

1. At the end of the pre-electrophoresis period.

TURN OFF THE POWER SUPPLY AND DISCONNECT BOTH

DC POWER CORDS FROM THE POWER SUPPLY. Open upper reservoir safety cover.

2. Immediately prior to loading the samples, rinse the wells of the gel thoroughly with electrophoresis buffer.

Use a loading tip or a syringe with a bent needle to wash away urea that has diffused into the wells.

4.3 Comb Volume/Maximum well volume

Note: To calculate sample well volume expressed in millimeters (mm) divide maximum volume by tooth depth.

-

Maximum Volume per tooth (microliters-ml)

# of wells

Tooth

width

0.75mm

thickness

1.0mm

thickness

1.5mm

thickness

100

3mm

22.5ml

30ml

45ml

Overall length of comb: 44.8cm

Tooth depth: 10mm
Spacing between teeth: 1.5mm

4.4 Electrophoresis

1. After loading the samples, attach the reservoir safety cover. Connect the DC power leads to the unit then to the

power supply with the proper polarity. Make sure the black leads are connected to the black cathodes (-) and

the red leads are connected to the red anode (+).

2. Turn on the power supply and set the voltage or wattage to the proper setting for the gel.

350V at 105mA/ gel

at 0.5X TBE buffer for 1.5mm thick gel.

3. Monitor the progress of electrophoresis by following migration of a marker dye front or by any other preferred

method.

4. When electrophoresis is complete,

TURN POWER SUPPLY OFF. Disconnect all DC power cords, first from

the power supply.

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