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5 / Nanopore coating protocol:
When working with biological samples, it is strongly recommended to use
the Izon reagent kit and coat nanopores. The Izon coating solution is used to
prevent the non-specific binding of protein from biological samples to the
nanopore. If left uncoated, the performance of nanopores may rapidly decline,
as the pore can become partially blocked and modified, and calibration
recordings need to be run more frequently. As the coating solution has a
greater viscosity than the electrolyte, the baseline may drift for 30 seconds or
so after coating.
— Replace the electrolyte in the lower fluid cell with freshly filtered coating
solution, fit the upper fluid cell and dispense 35
µ
L of coating solution into
the upper fluid cell.
— Fit the fluid cell cap, push in the PM2 nozzle and apply pressure of 20
mbar for 10 minutes. *
— Disconnect the PM2 nozzle, remove the upper fluid cell, pipette a droplet
of electrolyte on top of the nanopore, and replace the lower fluid with
electrolyte.
— Wipe away the electrolyte droplet, tap the upper fluid cell dry before
refitting it, 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.
*Use a 5 minute coating time for NP250-800s and 2 minutes for anything larger.
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