Shotgun Proteomics Sample Processing Automated by an Open-Source Lab Robot.

Shotgun Proteomics Sample Processing Automated by an Open-Source Lab Robot.
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DOI:
10.3791/63092
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发表时间:
2021-10-28
期刊:
Journal of visualized experiments : JoVE
影响因子:
--
通讯作者:
Lam MPY
Lam MPY
中科院分区:
其他
文献类型:
--
作者:
Han Y;Thomas CT;Wennersten SA;Lau E;Lam MPY

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Mass spectrometry-based shotgun proteomics experiments require multiple sample preparation steps, including enzymatic protein digestion and clean-up, which can take up significant person-hours of bench labor and present a source of batch-to-batch variability. Lab automation with pipetting robots can reduce manual work, maximize throughput, and increase research reproducibility. Still, the steep starting prices of standard automation stations make them unaffordable for many academic laboratories. This article describes a proteomics sample preparation workflow using an affordable, open-source automation system (The Opentrons OT-2), including instructions for setting up semi-automated protein reduction, alkylation, digestion, and clean-up steps; as well as accompanying open-source Python scripts to program the OT-2 system through its application programming interface.
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