Bioanalyzer

Enter the number of libraries to be analyzed in the chip above

Contact Nicole Wagner (new39@rutgers.edu) to schedule a bioanalyzer run and get instructions from her regarding how to give her a sample of your library.

The High Sensitivity Chip can handle 12 libraries up to 7 kilobases. If your library is larger, contact Nicole for pricing of a different chip.

Nicole does all of the steps below

Note that a Tapestation could also be used at this step.

Bioanalyzer Instructions

Prepping the Gel (use filtered gel within 4 weeks)

• Pipette 550uL gel matrix (red) into a spin filter
• Centrifuge at 1500g for 10 mins at room temp
• Aliquot 65uL filtered gel into 0.5mL RNase-free tubes

Prepping the Gel-Dye Mix

• Allow RNA dye (blue) to equilibrate to room temp for 30 mins (in the dark)
• Vortex RNA dye (blue) for 10 secs and spin down
• Add 1 uL RNA dye into a 65uL aliquot of filtered gel
• Vortex solution
• Spin gel-dye mix at 13,000g for 10 mins at room temp

Turn on Laptop/Bioanalyzer (use gel-dye mix within 1 day)

• Select correct icon
• Select correct program (RNA, DNA, protein)

Cleaning the Electrodes

• Slowly fill one of the wells of the electrode cleaner with 350uL RNase-free water
• Place the electrode cleaner in the bioanalyzer
• Close the lid and let sit for 5 mins
• Upon removing the electrode cleaner, keep lid open for at least 30 secs (to allow for evaporation)

Loading the Gel-Dye Mix

• Put a new chip on the chip priming station
• Pipette 9uL of gel-dye mix in the well marked G (white G on black background)
• Position the plunger at 1mL
• Close the chip priming station (MAKE SURE IT CLICKS)
• Press plunger down until it’s held by the clip
• Wait exactly 30 secs and release clip
• Wait exactly 5 secs and slowly pull plunger back to 1mL
• Open chip priming station and pipette 9uL gel-dye mix into wells marked G (black G on grey background)
• Discard remaining gel-dye mix

Loading the Conditioning Solution and Marker

• Pipette 9uL of RNA conditioning solution (white) into well marked CS
• Pipette 5uL of RNA marker (green) in all 11 sample wells and the ladder well

Loading the Diluted Ladder and Samples

• Pipette 1uL ladder into the well marked ladder
• Pipette 1uL of sample into each of the 11 sample wells (pipette 1uL RNA marker (green) into each unused sample well)
• Vortex chip for 1 min at 2400 rpm
• Run the chip in the bioanalyzer within 5 mins (RNA assay)

–CLEAN THE ELECTRODES AGAIN AFTER EVERY RUN–