Izon qNano, User Manual, Page: 39 / 56

The stretch value on the software display should now change as you adjust the
stretch manually.
2 / Nanopore wetting protocol
— Make up nanopore wetting solution according the Reagent Kit tech note.
— Add wetting solution to the lower (75
µ
L) and upper (35
µ
L) fluid cell
and fit the upper fluid cell.
— Fit the fluid cell cap, push in the PM2 nozzle and apply a pressure of 20
mbar for 4 minutes.* Apply a voltage of at least 0.1V so that during this
time a stable baseline can be observed being established.
If a baseline does not establish, disconnect the PM2 nozzle, remove the
fluid cell cap, and apply pulses of pressure with the Izon PAD up to 10
times. If this is still not successful, then try clicking the fluid cell cap by
putting pressure down on the nanopore arms and twisting until it clicks
into place.
— Remove the upper fluid cell, pipette a droplet of electrolyte on top of
the nanopore, and replace the lower fluid with electrolyte (prepared in
accordance with the Reagent Kit tech note).
— Wipe away the electrolyte droplet, fit the upper fluid cell and dispense
35
µ
L of electrolyte into the upper fluid cell.
— Fit the fluid cell cap and check the baseline current is stable, with an
RMS noise of <10 pA. Consult the troubleshooting guide at the end of this
manual if any difficulties are encountered at this stage.
* Use a 2 minute wetting time for NP250-800s and 30 seconds for anything larger.
To ensure that the pore is properly wetted, tap the fluid cell cap gently but
firmly a few times to dislodge any bubbles trapped in the aperture once the
baseline current has been initially established. The current should slowly
increase and reach a stable level once fully wetted.
After wetting, do not
click the fluid cell cap at all from this point and be very gentle when making
contact with the pore during pipetting.
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