Bio-Rad ReadyPrep™ 2-D Starter Kit - Manual (Page 18)

5 Using the forceps, carefully push the strip into the well as shown in Figure 13, taking care
not to trap any air bubbles beneath the strip. When pushing the IPG strips with the for-
ceps be certain the forceps are pushing on the plastic backing to the strip and not the gel
matrix.
6. Stand the gel(s) vertically by placing them in the gel box (Criterion system) or in a test tube
rack. Allow the agarose to solidify for 5 minutes before proceeding.
7. Mount the gel into the gel box per the instructions provided with the apparatus.
8. Fill the reservoirs with 1X Tris/glycine/SDS running buffer and begin the electrophoresis.
Table 4 lists the electrophoresis run conditions for each of Bio-Rad’s three SDS-PAGE for-
mats. The migration of the Bromophenol Blue, present in the overlay agarose solution, is
used to monitor the progress of the electrophoresis.
16
Fig. 13. Sliding ReadyStrip into IPG well filled with melted overlay agarose.
Strip Length 7 cm 11 cm 17 cm
Electrophoresis cell Mini-PROTEAN Criterion PROTEAN II XL
Conditions 200 V, constant 200 V, constant 16 mA/gel for
30 min, then 24
mA/gel for ~5 hr
Approximate run time 40 min 65 min 5.5 hr
Table 4.