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Shotgun Proteomics Sample Processing Automated by an Open-Source Lab Robot
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Biochemistry
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JoVE Journal Biochemistry
Shotgun Proteomics Sample Processing Automated by an Open-Source Lab Robot

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

DOI:
10.3791/63092-v

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10:12 min

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October 28, 2021

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, , , ,
1Department of Medicine-Cardiology,University of Colorado Anschutz Medical Campus, 2Department of Biochemistry & Molecular Genetics,University of Colorado Anschutz Medical Campus, 3Consortium for Fibrosis Research & Translation, School of Medicine,University of Colorado Anschutz Medical Campus

Chapters

  • 00:04Introduction
  • 00:40Mass Spectrometry (MS) Sample Preparation with a Single Protein
  • 04:44Peptide Clean-up Using SP3 Paramagnetic Beads
  • 06:19MS Sample Preparation with Protein Lysate
  • 08:37Results: Mass Spectrometry Proteomics with a Single Protein Standard Bovine Serum Albumin and a Detergent-Containing Human Heart Lysate Sample
  • 09:44Conclusion

Summary

Automatic Translation

Automatic Translation

Detailed protocol and three Python scripts are provided for operating an open-source robotic liquid handling system to perform semi-automated protein sample preparation for mass spectrometry experiments, covering detergent removal, protein digestion, and peptide desalting steps.

Tags

Keywords: Shotgun ProteomicsAutomated Sample ProcessingOpen-source Lab RobotLiquid Handling SystemMass SpectrometryProtein BiochemistryProteomic StudiesAmmonium BicarbonateDTTTemperature ControlAutomationOpen-source Python Scripts

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