Bio-Rad Experion RNA Analysis Kits - Manual (Page 15)

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If diluted RNA ladder is available from a previous preparation, skip steps 1 to 5. Do not reheat the
diluted ladder.
Start the run within 5 min of priming and loading the chip. For help with chip loading, refer to the
Experion Training Video: Chip Loading, available in the Experion software Help section under
Contents and Index > Contents > Appendices > Technical Videos.
3.4 Prepare the Samples and the RNA Ladder
Once the RNA samples and RNA ladder (red cap) have thawed on ice, vortex them briefly and spin
down for a few sec in a microcentrifuge. Both the sample and RNA ladder must be denatured. Use of
the RNA HighSens ladder also requires dilution.
3.4.1 Experion RNA StdSens Analysis
1. Pipet ≥2 μl RNA sample into separate RNase-free microcentrifuge tubes.
2. Pipet RNA ladder into an RNase-free microcentrifuge tube: use 1 μl RNA ladder for each chip plus an
extra 1 μl RNA ladder to accommodate variations in pipetting (for example, when running 1 chip, use
a total of 2 μl RNA ladder; for 3 chips, use 4 μl RNA ladder).
3. Denature the samples and the RNA ladder by heating the tubes at 70°C for 2 min. Place them on ice
for 2–5 min.
4. Vortex briefly and spin down. Keep the tubes on ice.
3.4.2 Experion RNA HighSens Analysis
1. Pipet 5 μl RNA ladder into an RNase-free microcentrifuge tube.
2. Transfer 2 μl RNA sample into separate RNase-free microcentrifuge tubes.
3. Denature the samples and the RNA ladder by heating the tubes at 70°C for 2 min. Place them on ice
for 2–5 min.
4. Spin down the ladder and samples in a microcentrifuge briefly and then place the tube on ice.
Add 795 μl DEPC-treated water to the denatured ladder. Keep the tubes on ice.
5. Aliquot diluted RNA ladder into RNase-free microcentrifuge tubes, store at –70°C, and avoid
exposing it to freeze-thaw cycles.
3.5 Prime the Chip
1. Pipet 9 µl GS into the highlighted well labeled GS (gel priming well, Figure 3.1).
2. On the priming station, set the pressure setting to B and the time setting to 1, as specified by the
alphanumeric code on the chip (Figure 3.1).
3. Open the Experion priming station and place the chip on the chip platform, matching the arrow on
the chip with the alignment arrow on the chip platform. A post on the chip prevents insertion in the
wrong position. Do not force the chip into position.
4. Close the priming station by pressing down on the lid. The lid should snap closed.
5. Press Start. A “Priming” message appears on the screen of the priming station, and the timer counts
down. Priming requires ~30 sec. Do not open the priming station during the countdown.
Experion RNA StdSens and HighSens Analysis Kits