70 Maintenance and Troubleshooting
6
Troubleshooting the RNA Application
Symptoms (RNA)
Residual Gel in Spin Filter after Centrifugation
Too High Quantitation Results
Back to “Symptoms (RNA)” on page 69
Most probable causes Solution
Gel was filtered at insufficient g-value. Refer to the Kit Guide for proper centrifuge settings.
Cooled centrifuge was used for gel
filtration.
Repeat centrifugation step at room temperature.
Gel was too cool or viscous. Reagents must be equilibrated at room temperature for 30 minutes prior to use.
Most probable causes Solution
Pipetting error during preparation of
ladder or samples.
Check dilution procedure and calibration of pipettes.
Chip pipetting error. Prepare new chip. Always insert the pipette tip to the bottom of the well when
dispensing the liquid. Use appropriate pipette and tips.
Insufficient vortexing of chip. Vortex chip for 1 minute. Only use the IKA vortexer. Adjust speed to set-point.
Probable causes Solution
RNA ladder degraded. Prepare a new chip using a new ladder aliquot. Always wear gloves when
handling chips and RNA samples to avoid contamination. Follow
decontamination procedure, see “How to Clean the Pin Set of the Electrode
Cartridge” on page 126.
Electrodes contaminated with RNases. Clean electrodes with RNaseZAP. Follow decontamination procedure, see “How
to Clean the Pin Set of the Electrode Cartridge” on page 126
Dye concentration too low. Use dye concentration according to the Kit Guide. Let the dye warm up to room
temperature for 30 min before preparing the gel-dye mix. Protect the dye from
light during this time.
RNA ladder not denatured. Heat denature the RNA ladder as described in the Kit Guide.
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