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Cell Biology JoVE (Open Access) Citable · DOI

Automated Robotic Liquid Handling Assembly of Modular DNA Devices

DOI: 10.3791/54703-v
What you'll learn
  • ✓ Configure modular DNA parts and generate liquid handler picklists using automated software
  • ✓ Execute combinatorial DNA device assembly on robotic liquid handling platforms
  • ✓ Verify clones and compare automated assembly efficiency to manual methods
Protocol

Here, an automated workflow to perform modular DNA "device" assembly using a modular cloning DNA assembly method on liquid-handling robots is presented. The protocol uses an intuitive software tool for generating liquid handler picklists for combinatorial DNA device library generation, which we demonstrate using two liquid handling platforms.

Difficulty
advanced
Total time
~2-3 days (including transformation, growth, and verification)
Biosafety
BSL-1

Steps

1
Specify DNA parts and generate liquid handler picklists

Use intuitive software to define modular DNA parts for library generation and automatically create picklists compatible with liquid handling robots.

▶ 00:50
2
Prepare plasmid DNA and assembly reagents

Prepare and quantify plasmid DNA and modular cloning reagents required for automated assembly reactions.

▶ 02:24
3
Execute DNA assembly on liquid handling robot

Load prepared reagents and DNA on the liquid handler and run the automated assembly script across multiple platforms.

▶ 03:55
4
Transform assembled reactions into bacterial cells

Perform chemical or electroporation transformation of automated assembly reactions into competent bacteria.

▶ 05:47
5
Verify clones by restriction digest and sequencing

Screen transformed clones using restriction analysis and DNA sequencing to confirm correct assembly.

▶ 07:56
6
Compare automated versus manual assembly outcomes

Analyze clone success rates and hands-on time for automated robotic assembly relative to manual preparation methods.

▶ 08:50
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