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Effects of DNA Template Preparation on Variability in Cell-Free Protein Production

Published

Author(s)

Eugenia Romantseva, Nina Alperovich, David Ross, Steven Lund, Elizabeth Strychalski

Abstract

DNA templates for protein expression remain an unexplored source of variability in the performance of cell-free expression (CFE) systems for protein production. To characterize this variability, we investigated the effects of two common DNA extraction methodologies, a postprocessing step, and manual versus automated preparation on protein production using CFE. We examined the effect of both the quantity and quality of the DNA template in the CFE reaction on the variability of protein produced. Based on these results, we offer practical guidance for preparing and characterizing DNA templates to achieve an acceptable amount of variability in CFE performance.
Citation
Synthetic Biology

Keywords

DNA template, cell-free expression, cell-free protein production, TX-TL, CFE, variability, reproducibility, automation

Citation

Romantseva, E. , Alperovich, N. , Ross, D. , Lund, S. and Strychalski, E. (2022), Effects of DNA Template Preparation on Variability in Cell-Free Protein Production, Synthetic Biology, [online], https://tsapps.nist.gov/publication/get_pdf.cfm?pub_id=933771 (Accessed October 10, 2025)

Issues

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Created August 13, 2022, Updated July 22, 2025
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