Bio-Rad Sequi-Gen GT Sequencing Cell - Manual (Page 17)

Fig. 4.5. Correct alignment of precision caster base with glass plate sandwich.
6. Lay the IPC assembly and attached precision caster base flat on a bench with the IPC
panel (drain port) facing up and the long edges of the clamps running parallel with the edge
of the benchtop.
• The most even pouring can be obtained by insuring that the assembly is level on the
benchtop. Failure to level the assembly may result in gel leakage. A leveling bubble
is provided to facilitate leveling the IPC assembly. Props (approximately 2 cm ) will
be required at the top of the IPC to level the unit for casting. The unit is now ready for
gel casting. An alternative to the use of props is to cast the gel with the precision caster
positioned off the edge of the lab bench.
Note: If casting a 38 x 50 cm IPC, place the 38 x 50 cm IPC assembly at an incline, with
the top of the apparatus approximately 4-5 cm higher than the bottom. (The bottom of
the apparatus contains the attached precision caster base). After the gel is cast, level the
assembly for gel polymerization.
7. While the gel solution is degassing, prepare a fresh 25% ammonium persulfate solution
(catalog number 161-0700).
• Choose the appropriate syringe and tubing assembly (tubing and luer taper) provided
with the precision caster. Insert the luer taper into the one end of the tubing. Secure the
other end of the tubing onto the luer end of the syringe.
8. When the gel solution has degassed, add 25% APS and TEMED (catalog number 161-0800)
in the recommended amounts (see Section 7.2).
• Swirl the solution gently to mix.
• Slowly pull the required gel volume into the syringe (see Table 4.1).
• Tap air bubbles to the top of the syringe (luer end) and gently force them out. If bub-
bles are inadvertently introduced into the tubing, pinch the portion of the tubing where
the bubbles exist while forcing some of the gel solution out. This should allow the
bubble to exit the tubing with the gel solution.
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