Bio-Rad ReadySub-Cell GT Cells - Manual (Page 17)

4.5 RNase Decontamination
Sub-Cell GT parts can be cleaned with a mild detergent and treated for 10 minutes with 3% hydrogen
peroxide (H
2
O
2
), and then rinsed with 0.1% DEPC- (diethyl pyrocarbonate) treated distilled water, to eliminate
RNases prior to using the Sub-Cell GT systems for RNA gels.1-2 Consult references 1 and 2 for other
suggestions regarding the use of DEPC in RNase decontamination.
Caution: DEPC is a suspected carcinogen. Always wear gloves, safety glasses, and a laboratory coat.
Use caution when handling DEPC-containing solutions. Consult the DEPC MSDS for more information.
Do not attempt to decontaminate RNase from Sub-Cell GT parts using extreme dry heat.
Note: Several commercial products are available for eliminating RNase contamination. RNaseZAP ® (Ambion)
is a safe, simple, and effective method that if used properly does not craze or fog the Sub-Cell GT parts. See
manufacturer’s instructions for proper use.
Section 5
Troubleshooting
Symptoms Cause Solutions
Slanted lanes (bands) Gel not fully solidified Let gel solidify for at least 30—45 min.
Comb warped or at an angle Check alignment of comb.
Curved line or distortion Bubbles in sample wells Remove bubbles prior to electrophoresis.
of lanes (bands)
Differential relative
Sample spilled out of wells Samples should have proper density.
mobilities Apply carefully.
Unit not leveled Level unit. Place on steady work bench.
Curved bands, smiles Sample overload Reduce load.
Ragged bands Sample density incorrect See sample application instructions.
Sample well deformed Carefully remove comb, especially from
soft gels. Be sure gel has solidified.
Cooling soft gels aids in comb removal.
Excessive power or heating Reduce voltage. See electrophoresis
instructions.
Band smearing and Agarose has improper Consult Bio-Rad about agarose.
streaking endosmosis (mr)
Salt concentration in Reduce salt concentration to ≤0.1 M.
sample too high
Excessive power and heating Reduce voltage. See electrophoresis
instructions.
Sample spilled out of well Apply sample carefully. Increase gel
thickness for large sample volumes.
Adjust comb height.
Incomplete digestion, Heat sample. Check enzyme activity.
nuclease contamination, Digest sample further.
bad enzyme
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