Automated DNA Normalization

Automating Normalization With The OT-2 (01:05)


Concentration normalization is a key component of many genomic and proteomic applications, such as NGS library prep and mass spectrometry sample prep. This process often requires working with 96 or more samples, all needing unique volumes of diluent to be brought to the correct “normalized” concentration. It’s a high-throughput and complex application, and can be cumbersome, labor-intensive, and error-prone to do by hand. This makes it ideal for automation.

A robot can more easily, quickly, and accurately read from a file and transfer the correct volume to each sample than a human. Automating this process means you don’t have to worry about errors in volume calculation or dispensing into the wrong location.

Using our OT-2 liquid handling robot and Normalization protocol, you can automate your normalization steps simply and efficiently.

Normalization can be used for the following applications:

  • Library Prep (Before and After)
  • DNA Normalization
  • Mass Spectrometry Sample Prep


Automating normalization offers many benefits to traditional manual protocols, such as:

  • Avoids human error
  • Increases speed of procedure
  • Assures maximum pipetting accuracy
  • Saves time, doing repetitive pipetting for you


Step 1

Place reagents onto robot

Load reagents and labware onto the deck of the OT-2.

Step 2

Create excel file

Create excel file indicating diluent volume for each well. Upload as CSV to our Protocol Library and download protocol.

Step 3

Calibrate and activate protocol

Confirm pipetting locations and hit “run.”

Step 4

Run your assay

Run your normalization protocol



The OT-2 supports various labware for normalization. While the labware you use may vary, we strongly recommend using one of our supported microplates and reservoirs with this protocol because that’s what we used to optimize and verify it. For more information on supported labware, please see our Labware Library.


Ensure that your OT-2 is fitted with single-channel pipettes for the volumes you will need to work with. Only single-channel pipettes are supported for this method. Please review our Opentrons Pipette White Papers linked below for more information on our pipettes and pipetting volumes as low as 1 µL.

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"I've done I don't know how many hundreds of minipreps in my life. I don't want to do another 3,000."
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