Bio-Rad Sequi-Gen GT Sequencing Cell - Manual (Page 25)

Section 5
Troubleshooting Guide
5.1 Operational Troubleshooter
The following table summarizes possible Sequi-Gen GT operational difficulties, probable
causes, and solutions. Refer to Sections 3 and 4 for recommended procedures.
Problem Probable Causes Solution
No gel solution entering gel mold
Gel solution is leaking into the
precision caster base
Gel solution is receding from
the top of the gel
Bubbles in gel
• Luer taper, tubing or syringe
orifices blocked.
• Precision Caster Base injection
port misaligned with gap
between the glass plates
• Precision caster gasket missing
• Gasket hole not aligned with the
injection port of the precision
caster base
• Glass plates and gasket are not
in contact
• Glass plates, spacers and
clamps are not flush at their
bottom edge
• Entire casting assembly (preci-
sion caster assembly and IPC
assembly) at too high of an
incline
• Glass plates, spacers and
clamps are not flush
• Entire casting assembly (preci-
sion caster assembly and IPC
assembly) at an incline
• Air bubbles injected into the
mold because gel was injected
too quickly
• Gel solution not degassed
• Air trapped in tubing
• Air bubbles trapped in syringe
• Air injected into the gel mold
because gel volume was
inadequate
• Thoroughly clean syringe,
tubing and tapers of dried gel
solution with warm water and
mild detergent (see Section 3)
• Readjust precision caster base
so that white slit can be seen
between the green glass plates
while looking through the
injection port (see section 4)
• Insert gasket into base
• Disassemble precision caster
base from the IPC assembly,
adjust the gasket and reassemble
• Disassemble and make sure the
bottom edges of the clamps,
glass plates and spacers are
flush
• Disassemble and make sure the
bottom edges of the clamps,
glass plates and spacers are flush
• Lower the casting assembly or
completely level the assembly
• Disassemble and make sure the
bottom edges of the clamps,
glass plates and spacers are flush
• Level the casting assembly
• Refer to injection rate recom-
mendations under Note on Gel
Bubble Formation, Section 4.3
• Degas gel before casting
• Remove tubing before drawing
gel solution into the syringe,
then attach tubing and gently
push gel solution through tubing
• Draw gel solution into the
syringe barrel slowly to avoid
introducing bubbles on the side
of the barrel
• See Section 4.4 (Table 4.2) for
suggested gel volumes
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