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Enhancing Yeast Transformation: Achieving a 10-Fold Increase through a Single Adjustment in the Lithium Acetate-Polyethylene Glycol Method
  • +2
  • Mathilde Kadouch,
  • Pierre Gaspin,
  • Christelle Marchal,
  • Sabine Castano,
  • Christophe Cullin
Mathilde Kadouch
Universite de Bordeaux - Pessac Campus
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Pierre Gaspin
Universite de Bordeaux - Pessac Campus
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Christelle Marchal
Universite de Bordeaux - Pessac Campus
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Sabine Castano
Universite de Bordeaux - Pessac Campus
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Christophe Cullin
Universite de Bordeaux - Pessac Campus

Corresponding Author:[email protected]

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Abstract

The Lithium-PEG method to transform yeast cells is a common experiment used by most yeast laboratories. After several optimizations, this method can yield up to 10 6 transformants / µg of plasmid. Some experiments such as library screening or complex transformation require to maximize transformation yield. Here we show that the addition of a 2M sorbitol solution acts as an osmo-protectant during and after heat shock and increases ten-fold the transformation rate. This optimization requires only one additional pipetting step compared to the initial protocol, and is therefore practical for a daily use.
11 Sep 2024Submitted to Yeast
12 Sep 2024Submission Checks Completed
12 Sep 2024Assigned to Editor
12 Sep 2024Review(s) Completed, Editorial Evaluation Pending
17 Sep 2024Reviewer(s) Assigned
30 Sep 2024Editorial Decision: Revise Major
14 Nov 20241st Revision Received
18 Nov 2024Submission Checks Completed
18 Nov 2024Assigned to Editor
18 Nov 2024Review(s) Completed, Editorial Evaluation Pending
18 Nov 2024Reviewer(s) Assigned