Home Cell Biology Automated Protocols for Macromolecular Crystallization at the MRC Laboratory of Molecular Biology
Cell Biology JoVE (Open Access) Citable · DOI

Automated Protocols for Macromolecular Crystallization at the MRC Laboratory of Molecular Biology

DOI: 10.3791/55790-v
What you'll learn
  • Operate automated systems for high-throughput crystallization plate preparation
  • Set up nanoliter-scale vapor diffusion droplets using robotic dispensing
  • Screen macromolecular crystallization conditions systematically
Protocol

Automated systems and protocols for the routine preparation of a large number of screens and nanoliter crystallization droplets for vapor diffusion experiments are described and discussed.

Difficulty
advanced
Total time
~2–4 hours per screening experiment (depending on plate count and sample number)

Steps

1
Prepare 72 crystallization plates with screening kits

Use automated System 1 to fill 72 crystallization plates with commercial screening reagents. This automated preparation ensures consistent reagent distribution across multiple plates for routine high-throughput screening.

▶ 00:59
2
Set up 100 nL crystallization droplets in 96-well plates

Deploy automated System 2 to dispense 100 nanoliter crystallization droplets from a single macromolecular sample into 20 96-well plates. This nanoliter-scale setup enables efficient vapor diffusion screening across numerous conditions.

▶ 04:46
3
Screen outer membrane protein F crystallization conditions

Evaluate initial crystallization results from OmpF and MreB samples using the automated screening outputs. This demonstrates how the automated protocols identify promising crystallization conditions for further optimization.

▶ 09:22
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