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Protein Dynamics in Living Cells

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Related Experiment Video

Updated: Sep 12, 2025

A Robotic Platform for High-throughput Protoplast Isolation and Transformation

A Robotic Platform for High-throughput Protoplast Isolation and Transformation

Published on: September 27, 2016

Glow with the Flow: Reproducible Analysis of Transiently Transformed Protoplasts Using Dual Fluorescent Reporters in

Joseph S Taylor¹, Elyse A Shoppell², Bastiaan O R Bargmann¹

¹School of Plant and Environmental Sciences, Virginia Tech, Blacksburg, VA, USA.

Biorxiv : the Preprint Server for Biology

|August 6, 2025

Summary

This summary is machine-generated.

This study introduces an open-source R workflow to automate positive fluorescent selection in plant protoplast transformation assays. This enhances the reproducibility and scalability of gene regulation studies.

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Last Updated: Sep 12, 2025

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Area of Science:

Plant Biology
Molecular Biology
Bioinformatics

Background:

Transient transformation assays using protoplasts are common in plant research.
Positive fluorescent selection with flow cytometry aids in analyzing transfected cells.
Current methods require significant manual effort and subjective analysis.

Purpose of the Study:

To develop an automated, reproducible, and scalable workflow for analyzing plant protoplast transient transformation.
To improve the quantification of reporter gene expression in successfully transfected plant cells.

Main Methods:

Developed an open-source workflow using the R programming language.
Integrated flow cytometry data analysis for positive fluorescent selection.
Automated quantification of reporter gene expression in transfected protoplasts.

Main Results:

The R workflow streamlines the analysis of positive fluorescent selection data.
Enhanced reproducibility and scalability of transient transformation assays.
Reduced manual effort and subjective judgment in data interpretation.

Conclusions:

The new R workflow offers a robust solution for analyzing plant protoplast transformation experiments.
Facilitates rapid and reliable study of gene regulation and signal transduction in plants.
The open-source workflow promotes wider adoption and collaboration in plant synthetic biology.