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AUTOMATED OPENTRONS LOADING PROTOCOL WITH EVOTIP PURE

Complete, automated end-to-end sample loading on the Opentrons OT-2 through a layered sandwich approach with defined airgaps between the layers. The sandwich is pushed through the Evotip with the OT-2 pipette for 100 seconds leaving the sample safely stored and ready for injection on the Evosep One.

The automated sample loading protocol sequentially deposits 15 µl solvent A, 20 µl sample, and 150 µl solvent A to facilitate equilibration, loading, and desalting followed by storage, respectively.

On-robot Evotip loading requires an Evotip Loading kit (OT-2) (EV1144) and allows for full end-to-end sample processing on the Opentrons without manual intervention.

Stand-alone automated Evotip loading has been converted into an easy-to-use HTML form, that generates a complete python script for use in the Opentrons app, allowing the user to load from 8 to 288 Evotips in a single run.

END-TO-END SAMPLE PREPARATION WITH OT-2 PROTEIN ON-BEAD DIGESTION AND EVOTIP LOADING

Three end-to-end workflows utilizing protein aggregation capture and Evotip loading. They all contain steps with aggregation-based protein capture on magnetic beads, clean-up, and digestion as well as seamless Evotip loading based on layered sandwich approach.

The protocols have been converted into an easy-to-use HTML form, that generates a complete python script to use in the Opentrons app. It allows the user to process from 8 to 192 samples in a single run.

One workflow is designed to handle any type of reduced and alkylated cell lysate. Two workflows are designed for the analysis of unprocessed plasma. One generates samples for the analysis of neat plasma, while the other utilizes Mag-Net enrichment for deep plasma proteomics.

Protocols

Evosep assumes no responsibility and shall have no liability for any damage or loss of samples, material and hardware that may arise from use of or in connection with Evosep recommended protocols and SOP’s.

OPENTRONS OT-2 NEAT PLASMA AND EVOTIP LOADING PROTOCOL

The Opentrons OT-2 neat plasma and Evotip loading protocol allows for a fully automated, end-to-end sample preparation process from raw plasma. This protocol seamlessly integrates the sample preparation and Evotip loading steps into one workflow. Utilizing the Opentrons 8-channel P300 pipettes, it can efficiently handle up to 192 samples per run.

Generate protocol file and import to Opentrons app.

OPENTRONS OT-2 MAG-NET PLASMA AND EVOTIP LOADING PROTOCOL

The Opentrons OT-2 Mag-Net plasma and Evotip loading protocol allows for fully automated, end-to-end deep plasma proteomics from 4 µl raw plasma. This protocol seamlessly integrates the sample preparation and Evotip loading steps into one workflow. Utilizing the Opentrons 8-channel P300 pipettes, it can efficiently handle up to 192 samples per run.

Generate protocol file and import to Opentrons app.

Opentrons OT-2 PAC digestion protocol

The Opentrons OT-2 protein aggregation capture (PAC) protocol and Evotip loading protocol allows for fully automated, end-to-end analysis of reduced and alkylated protein lysates. This protocol seamlessly integrates the sample preparation and Evotip loading steps into one workflow. Utilizing the Opentrons 8-channel P300 pipettes, it can efficiently handle up to 192 samples per run.

Generate protocol file and import to Opentrons app.

OPENTRONS OT-2 Automated loading protocol

The Opentrons OT-2 automated loading protocol allows for simultaneous concentration, desalting, and storage of peptide samples on Evotips. This protocol allows for completely automated, hands-free loading of up to 288 samples in one run with the Opentrons P300 and P20 pipettes.

Generate protocol file and import to Opentrons app.

Step-by-Step Guide

Opentrons ot-2 Evotip Loading protocol

Download the step-by-step guide on how to use the Opentrons OT-2 Evotip Loading Protocol.

See video guides below as well.

Opentrons OT-2 Protein on-bead digestion and Evotip Loading Protocol

Download the step-by-step guide on how to use the Opentrons OT-2 Protein On-Bead Digestion and Evotip Loading Protocol.

Application notes

Cost-efficient and scalable end-to-end workflow on the Opentrons OT-2 for neat plasma utilizing Evotip Pure

App note describing end-to-end workflow on the Opentrons OT-2 for neat plasma.

A complete, automated Opentrons OT-2 loading protocol for simplified sample loading of Evotip Pure

App note describing automatic loading protocol for loading of Evotips.

End-to-end, fully automated digestion protocol and Evotip Pure workflow on the Opentrons OT-2

App note describing digestion and automatic loading protocol for loading of Evotips.