StarLab StarPhoresis N2510-1010 Manuel D'utilisation page 8

TABLE C: Approximate gel solution volumes
Gel solution volume,
0.8mm spacer thickness
Gel solution volume,
1.5mm spacer thickness
2. Run the acrylamide gel solution mix slowly down the inside edge of the gel cassette. Avoid aeration.
Place the comb in the gel plate assembly.
If a stacking gel is to be used, carefully overlay the gel solution to a depth of 3
buffer or water-saturated butanol. Following polymerization of the separating gel, pour off the overlay
layer (rinse off butanol with electrophoresis gel buffer) and pour a stacking gel if required. Insert the
comb ensuring bubbles are not trapped around comb teeth. Once the stacking gel has polymerised
use the gel immediately or store wrapped in a damp paper towel and plastic film at 4Cº. Wait a mini-
mum of 15 minutes for the gel to polymerise. Repeat process as required.
3. Release the cams and pull away from the upper buffer chamber and gels. Wash off any residual acry-
lamide. Place the upper buffer chamber into the lower buffer chamber. Stainless steel pins are located
on the lower sides of the upper buffer chamber that slide into the precision machined clear sides of
the lower buffer chamber to set it in place.
4. Add the appropriate volume of running buffer to the upper buffer chamber (Table B gives approxi-
mate volumes), making sure the running buffer is 3mm below the top of the blank glass, ensuring
sufficient contact with the top of the gel surface. Be sure that the running buffer is not leaking from
the upper buffer chamber to the lower buffer chamber. If buffer is leaking you will need to drain the
UBC and reset the gel cassettes.
NOTE: When running only one gel a blocking plate is required on the other side of the unit to retain
the top buffer level.
Gel and buffer volumes
Some guidelines for general operating conditions are given in Table B, but conditions vary according to the
number of gels, their composition, length, and cross sectional area. The current requirement will increase in
proportion to the number of gels or gel thickness providing that the voltage is not limiting, eg. two gels
require twice the current of one, but the same voltage. Longer gels require proportionally higher voltages.
By increasing the gel concentration the electrical resistance is increased and the rate of migration decreases.
Higher voltages can be applied but be careful not to overheat the gel. The conductivity of non-dissociating
buffer systems gels vary enormously and conditions must be determined empirically.
The run conditions are to be taken as a guideline only and apply to SDS Tris-glycine gels. If the plates
become hot increase the water flow rates within the recommended limits or reduce the power settings.
STARLAB offers a broad range of power supplies for a number of electrophoresis separations.
Sample loading
• If a native gel is being used, pre-electrophorese the gel for 15
SDS gels do not need this step.
• Centrifuge samples at 12,000xg for 5 minutes. If this step is omitted samples may streak during
electrophoresis.
• Carefully remove the sample comb and immediately flush the wells with electrophoresis buffer using a
syringe.
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StarPhoresis 2-Gel Mini and Wide Mini Vertical Electrophoresis Systems
N2510-1010 N2516-1410
7.5ml
15ml
N2520-2010
13.5ml 24.6ml
27ml 49.1ml
40 minutes prior to loading samples.
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N2520-1010
15ml
30ml
5mm with 1 x gel
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