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Automated Platform for the Plasmid Construction Process

  • Alberto A. Nava
  • , Anna Lisa Fear
  • , Namil Lee
  • , Peter Mellinger
  • , Guangxu Lan
  • , Joshua McCauley
  • , Stephen Tan
  • , Nurgul Kaplan
  • , Garima Goyal
  • , R. Cameron Coates
  • , Jacob Roberts
  • , Zahmiria Johnson
  • , Romina Hu
  • , Bryan Wu
  • , Jared Ahn
  • , Woojoo E. Kim
  • , Yao Wan
  • , Kevin Yin
  • , Nathan Hillson
  • , Robert W. Haushalter
  • Jay D. Keasling*
*Corresponding author for this work
    • Joint Bioenergy Institute
    • Lawrence Berkeley National Laboratory
    • University of California at Berkeley

    Research output: Contribution to journalJournal articleResearchpeer-review

    267 Downloads (Orbit)

    Abstract

    There is a growing need for applications capable of handling large synthesis biology experiments. At the core of synthetic biology is the process of cloning and manipulating DNA as plasmids. Here, we report the development of an application named DNAda capable of writing automation instructions for any given DNA construct design generated by the J5 DNA assembly program. We also describe the automation pipeline and several useful features. The pipeline is particularly useful for the construction of combinatorial DNA assemblies. Furthermore, we demonstrate the platform by constructing a library of polyketide synthase parts, which includes 120 plasmids ranging in size from 7 to 14 kb from 4 to 7 DNA fragments.

    Original languageEnglish
    JournalACS Synthetic Biology
    Volume12
    Issue number12
    Pages (from-to)3506-3513
    ISSN2161-5063
    DOIs
    Publication statusPublished - 2023

    Keywords

    • Synthetic biology
    • Automation
    • Plasmid construction
    • Polyketide synthases

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