Bio-Rad Experion RNA Analysis Kits - Manual (Page 14)

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3.1 Set Up the Electrophoresis Station
1. If needed, perform a deep cleaning of the electrodes (see Appendix B for instructions).
2. Power on the computer and then power on the Experion electrophoresis station by pushing the
green button in the center of the front panel. The steady green LED above the button indicates the
unit is on.
3. Launch Experion software. If the instrument and computer are communicating properly:
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A green dot and the last 4 digits of the instrument serial number appear at the lower right of the
software screen
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The electrophoresis station icon appears in the upper left corner
When there is no connection, these indicators are absent and a grayed-out instrument icon appears
at the upper left of the software screen.
3.2 Equilibrate the Kit Reagents
1. Set a heating block or water bath to 70°C. You will use this heating device to denature the samples
and the RNA ladder later in the protocol.
2. Thaw the following kit components on ice (15–20 min):
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RNA samples
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RNA ladder (red cap)
3. Equilibrate the following kit components to room temperature (15–20 min):
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RNA stain (blue cap)
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RNA loading buffer (yellow cap)
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RNA gel (green cap) (if filtered gel was prepared previously, remove it from storage and
equilibrate to room temperature)
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Sensitivity enhancer (clear cap, RNA HighSens only)
4. Invert each tube several times and then vortex to reincorporate any condensed liquid. Briefly
centrifuge the solutions to the bottom of the tubes. Make sure the RNA stain solution (blue cap) is
thawed before proceeding.
3.3 Filter the Gel and Prepare the Gel-Stain Solution
A gel-stain solution (GS) preparation is sufficient for use with up to three RNA chips. Prepare
GS daily and keep it covered with foil until ready for use. If filtered gel (G) is available, skip step 1.
Use G within 1 month of preparation. After 1 month, refilter it before reuse.
1. Pipet 600 μl RNA gel (green cap) into a spin filter and centrifuge it at 1,500 x g for 10 min. Inspect the
tube to ensure all of the gel has passed through the filter and then discard the filter.
2. Prepare the GS. Pipet 65 μl G into an RNase-free 0.65 ml microcentrifuge tube, add 1 μl RNA stain,
and vortex for 10 sec. Wrap the tube of GS in aluminum foil to protect the stain from light.
3. RNA HighSens analysis only: Centrifuge the GS at 13,000 x g for 10 min.
Experion Automated Electrophoresis System