Bio-Rad Sequi-Gen GT Sequencing Cell - Manual (Page 23)

Table 4.3 (continued)
Sequi-Gen GT Gel Recommended
Cell Size Thickness Power Setting
38 x 30 cm 0.25 mm 70-75 W
38 x 30 cm 0.40 mm 70-75 W
38 x 30 cm 0.75 mm 70-75 W
38 x 30 cm 0.25-0.75 mm 70-75 W
38 x 30 cm 0.40-1.20 mm 70-75 W
38 x 50 cm 0.25 mm 70-80 W
38 x 50 cm 0.40 mm 75-85 W
38 x 50 cm 0.75 mm 75-85 W
38 x 50 cm 0.25-0.75 mm wedge 75-85 W
38 x 50 cm 0.40-1.2 mm wedge 75-85 W
Important: Never allow the gel temperature to exceed 60 °C. Severe damage to the glass
or adhesive bond may result.
Caution: Periodically check the level of the upper buffer to make sure that it is at least
1 cm above the short glass plate.
2. Continue gel electrophoresis until the desired fragment size separation is achieved. Typically,
gel electrophoresis times are monitored by observing the dye front mobility of either the
bromophenol blue (“fast blue”) or xylene cyanol (“slow blue”) during the course of elec-
trophoresis. Fragment and dye front mobility as a function of polyacrylamide percentage are
shown in Table 4.4 below, and should be used as a guide for gel electrophoresis monitoring.
Table 4.4 Migration of Single-stranded DNA in Denaturing Polyacrylamide
Gels in Relation to Dye Marker Gel Migration*
Polyacrylamide Bromophenol
Gel Percentage Blue Xylene Cyanol
5% 35 bases 130 bases
6% 26 bases 106 bases
8% 19 bases 75 bases
10% 12 bases 55 bases
* From Ausubel, F. M. et. al., Current Protocols in Molecular Biology, Greene and Wiley, 1993.
4.7 Disassembly
1. When the desired dye front mobility has been achieved, turn off the power supply, and
remove both safety covers.
• The upper buffer chamber can be partially emptied by inserting the drain port
connector (and any attached tubing) into the drain port on the IPC. A “click” will be
heard when the drain port/tubing connector has been properly inserted (Figure 4.10).
• Buffer will begin to drain from the IPC immediately after the connector is inserted
into the drain port.
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