Bioanalyzer RNA 6000 Nano Kit
1.
Preparing the gel
1.1.
Pipette 500ul of RNA 6000 Nano gel mix into spin filter.
1.2.
Centrifuge at 8000 rpm for 10 minutes at room temperature.
1.3.
Use filtered gel within 4 weeks.
2.
Preparing the Gel-Dye mix
2.1.
Allow the RNA 6000 Nano dye concentrate to equilibrate to room temperature for 30 minutes.
2.2.
Vortex RNA 6000 Nano dye concentrate for 10s, spin down briefly and add 1ul dye to
65ul aliquot of filtered gel (sufficient to run 2 chips).
2.3.
Vortex the solution well. Spin tube at 13,000rpm for 10 minutes at room temperature.
3.
Preparing samples
3.1.
Aliquot 3ul of each sample into a PCR tube.
3.2.
Obtain an aliquot of ladder for each chip to be run.
3.3.
Denature the ladder and samples for 2 minutes at 70 o C in a PCR machine.
3.4.
Snap freeze on ice.
3.5.
Pulse spin in a cooled centrifuge.
4.
Loading the gel dye mix
4.1.
Put a new RNA 6000 Nano chip on the chip priming station.
4.2.
Pipette 9ul of gel-dye mix into well marked – G in black circle.
4.3.
Close chip priming station.
4.4.
Press plunger until its is held by the clip.
4.5.
Wait for exactly 30s the release clip.
4.6.
Pipette 9ul of gel mix into the two wells marked – G in grey box.
5.
Loading the Agilent 6000 Nano marker
5.1.
Pipette 5ul of RNA 6000 nano marker into all 12 sample wells and the well marked ladder.
6.
Loading the ladder and samples.
6.1.
Pipette 1ul of the prepared ladder in the well marked ladder.
6.2.
Pipette 1ul of each sample into the 12 sample wells (pipette 1ul of RNA 6000 nano marker into any unused wells.
6.3.
Put the chip horizontally in the adapter and vortex for 1 minute at 2400rpm.
6.4.
Run the chip in the Agilent 2100 Bioanalyzer within 5 minutes.

7.
Setting up the bioanalyzer
7.1.
Switch on the bioanalyzer at the back of the machine.
7.2.
Switch on the computer.
7.3.
User name: Administrator, Password: 3000hanover
7.4.
Select 2100 Expert
7.5.
Clean electrodes:
7.5.1.
1 minute with 2x200ul RNase ZAP
7.5.2.
10s with 2x200ul DEPC water
7.5.3.
Air dry for 30s, close lid.
7.6.
Select Assay > Electrolysis > RNA > Eukaryotic Total RNA Nanoseries II.
7.7.
Press Start.