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Related Experiment Video

Updated: Nov 29, 2025

Automated Robotic Liquid Handling Assembly of Modular DNA Devices

Automated Robotic Liquid Handling Assembly of Modular DNA Devices

Published on: December 1, 2017

Automating Cloning by Natural Transformation.

Xinglin Jiang¹, Emilia Palazzotto¹, Ewa Wybraniec²

¹The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, 2800 Kgs. Lyngby, Denmark.

ACS Synthetic Biology

|November 24, 2020

Summary

This study introduces automatic natural transformation (ANT) cloning, a simple and affordable method for DNA cloning and engineering. ANT bypasses traditional bottlenecks, enabling high-throughput synthetic biology applications.

Keywords:

Acinetobacter baylyi ADP1 automated cloning benchtop robot biosynthetic gene clusters natural transformation

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Last Updated: Nov 29, 2025

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A Robotic Platform for High-throughput Protoplast Isolation and Transformation

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Published on: September 27, 2016

Area of Science:

Synthetic Biology
Molecular Biology
Microbiology

Background:

Traditional DNA cloning methods, such as those using *E. coli*, often require complex procedures like heat shock or electroporation, posing a bottleneck for automated workflows in synthetic biology.
Developing accessible and automated cloning platforms is crucial for advancing synthetic biology research and applications.

Purpose of the Study:

To develop a simplified, automated, and cost-effective DNA cloning and engineering platform for synthetic biology.
To overcome the limitations of traditional cloning methods by exploring bacterial natural transformation.

Main Methods:

Explored bacterial natural transformation in *Acinetobacter baylyi* ADP1 for direct cloning of DNA fragments generated by Gibson assembly or overlap extension PCR (OE-PCR).
Developed an automatic natural transformation (ANT) cloning protocol that eliminates the need for DNA purification, competence induction, or specialized equipment.
Validated the ANT method by cloning and engineering 21 biosynthetic gene clusters (BGCs) of varying sizes and GC content.

Main Results:

Achieved efficient recombinant plasmid generation directly from PCR products in *A. baylyi* ADP1 cultures.
Obtained high colony yields (up to 10,000 colonies per microgram of DNA) with a low rate of false positive colonies.
Successfully cloned and engineered diverse BGCs ranging from 1.5 to 19 kb, including a nucleoside BGC exhibiting antibacterial activity.
Demonstrated consistent cloning efficiency when the ANT method was adapted to a low-cost benchtop robot.

Conclusions:

Automatic natural transformation (ANT) cloning offers a robust, user-friendly, and affordable alternative for high-throughput DNA engineering.
This method significantly simplifies the cloning process, making advanced synthetic biology techniques more accessible.
ANT facilitates rapid engineering of complex genetic constructs, accelerating discovery in areas like natural product biosynthesis.