This video will show you how to wash a flow cell after a sequencing run and how to load a new library. We strongly recommend all our users to view the demonstration video before performing this method:

Washing and reloading a MinION and GridION Flow Cell video

Mix well by pipetting, and place the tube(s) on ice.

Note: Do not vortex the Flow Cell Wash Mix tube.

Please ensure your flow cell is not sequencing while performing any of the wash steps. Manipulating the flow cell while it is sequencing can results in damage and pore loss.

Note: It is vital that the flow cell priming port and SpotON sample port are closed before removing the waste buffer to prevent air from being drawn across the sensor array area, which would lead to a significant loss of sequencing channels.

You should remove the full volume of waste buffer from the waste channel. If required you can dispense the volume aspirated from the waste channel and repeat the process to remove the full volume from the waste channel.

1. Set your P1000 pipette to 1000 µl.
2. Insert the P1000 pipette tip into waste port 1.
3. Slowly aspirate to remove the waste buffer from waste port 1.

Note: As both the priming port and SpotON sample port are closed, no fluid should leave the sensor array area.

1. Set a P1000 pipette to 200 µl.
2. Insert the tip into the flow cell priming port.
3. Turn the wheel until the dial shows 220-230 µl, or until you can see a small volume of buffer/liquid entering the pipette tip.
4. Visually check that there is continuous buffer from the flow cell priming port across the sensor array.

1. Using a P1000 pipette, take 200 µl of the flow cell wash mix
2. Insert the pipette tip into the priming port, ensuring there are no bubbles in the tip
3. Slowly twist the pipette wheel down to load the flow cell (if possible with your pipette) or push down the plunger very slowly, leaving a small volume of buffer in the pipette tip.

Note: Dispensing the wash mix into your flow cell should take at least 5 seconds, ensuring it is done very slowly and at a continuous rate.

1. Set a P1000 pipette to 200 µl
2. Insert the tip into the priming port
3. Turn the wheel until the dial shows 220-230 µl, to draw back 20-30 µl, or until you can see a small volume of buffer entering the pipette tip
   

Note: Take care when drawing back buffer from the flow cell, do not remove more than 20-30 µl. Visually check that there is continuous buffer from the priming port across the sensor array.

1. Using a P1000 pipette, take 800 µl of the priming mix.
2. Insert the pipette tip into the priming port, ensuring there are no bubbles in the tip.
3. Slowly twist the pipette wheel down to load the flow cell (if possible with your pipette) or push down the plunger very slowly, as illustrated in the demonstration video, leaving a small volume of buffer in the pipette tip.

Note: Dispensing the 800 µl of priming mix into your flow cell should take at least 10 seconds, ensuring it is done very slowly and at a continuous rate.

During this time, prepare the library for loading by following the steps below.

You should remove the full volume of waste buffer from the waste channel. If required you can dispense the volume aspirated from the waste channel and repeat the process to remove the full volume from the waste channel.

1. Set your P1000 pipette to 1000 µl.
2. Insert the P1000 pipette tip into waste port 1.
3. Slowly aspirate to remove the waste buffer from waste port 1.

Note: As both the priming port and SpotON sample port are closed, no fluid should leave the sensor array area.

1. Open the priming port and gently lift open the SpotON sample port.
2. Using a P1000 pipette, take 200 µl of the priming mix.
3. Insert the pipette tip into the priming port, ensuring there are no bubbles in the tip.
4. Slowly twist the pipette wheel down to load the flow cell (if possible with your pipette) or push down the plunger very slowly, as illustrated in the demonstration video, leaving a small volume of buffer in the pipette tip.

Note: Dispensing the 200 µl of priming mix into your flow cell should take at least 5 seconds, ensuring it is done very slowly and at a continuous rate.

1. Carefully place the leading edge of the light shield against the clip. Note: Do not force the light shield underneath the clip.

2. Gently lower the light shield onto the flow cell. The light shield should sit around the SpotON cover, covering the entire top section of the flow cell.

1. Set a P1000 pipette to 200 µl
2. Insert the tip into the priming port
3. Turn the wheel until the dial shows 220-230 µl, to draw back 20-30 µl, or until you can see a small volume of buffer entering the pipette tip
   

Note: Take care when drawing back buffer from the flow cell, do not remove more than 20-30 µl. Visually check that there is continuous buffer from the priming port across the sensor array.

1. Using a P1000 pipette, take 500 µl of the Storage Buffer (S).
2. Insert the pipette tip into the priming port, ensuring there are no bubbles in the tip
3. Slowly twist the pipette wheel down to load the flow cell (if possible with your pipette) or push down the plunger very slowly, leaving a small volume of buffer in the pipette tip.

Note: Dispensing the Storage Buffer (S) into your flow cell should take at least 5 seconds, ensuring it is done very slowly and at a continuous rate.

You should remove the full volume of waste buffer from the waste channel. If required you can dispense the volume aspirated from the waste channel and repeat the process to remove the full volume from the waste channel.

1. Close the priming port.
2. Set your P1000 pipette to 1000 µl.
3. Insert the P1000 pipette tip into waste port 1.
4. Slowly aspirate to remove the waste buffer from waste port 1.

Note: As both the priming port and SpotON sample port are closed, no fluid should leave the sensor array area.