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5. Sequencing run
  • Introduction
  • 1. Software installation
  • 2. Hardware check
  • 3. Flow cell check
  • 4. Library preparation
  • 5. Sequencing run
  • 6. Results and metrics
  • 7. Next steps
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You are in the ‘Windows’ branch of Software installation.
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5.5

Load your flow cell

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Important

Pipetting tips

The following section will load fluid and sample into the flow cell. In the images below are some simple tips on pipetting technique which are essential to avoid introducing air bubbles and damaging the sensor array.

When drawing up solution:

  • make sure there is no air plug at the bottom of the tip

When ejecting the solution:

  • do not fully expel the liquid from the tip
  • leave a small volume in the tip end so that no air can follow the solution into the inlet port

pipette-tips3

Important

The following steps will explain how to add your sample to your flow cell.

They should be done in quick succession to ensure correct operation of the product.

Please keep your MinION Mk1B flat, espescially when the sample and priming ports are open.

Open the priming port by sliding the cover clockwise so that the port is visible.

open-priming-port

Open the sample port by gently lifting the sample port cover. You will still need to use the priming port, so keep this open.

5.5.3

Tip

On the flow cell, the sample port is labelled SpotON.

Using a P1000 pipette, slowly load 200 µl of the Priming Mix into the priming port.
Do NOT eject to the last stop of the pipette, as this would introduce air bubbles.
Pipette slowly, to avoid the buffer bubbling out of the sample port.
Carry out this step immediately before loading the library.

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Resuspend the Pre-Sequencing Mix (Tube 5) by gently pipetting it up and down using a P200 pipette and tip, to get a homogenous mixture.

ligv2-seq3

Tip

In the next step, pipette up and down carefully to avoid creating air bubbles. Gentle resuspension also helps prevent accidental shearing of the DNA.

Using a P200 pipette, hold the pipette tip just above the sample port and slowly load 75 µl, drop by drop, of the Pre-Sequencing Mix (Tube 5).

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  • Close the sample port with the cover ensuring the bung enters into the port
  • Close the priming port
  • Close the MinION Mk1B lid

5.5.9

Prev 5.6 Start your run

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