Effects of DNA template preparation on variability in cell-free protein production

Autor:	Eugenia Romantseva, Nina Alperovich, David Ross, Steven P Lund, Elizabeth A Strychalski
Rok vydání:	2022
Předmět:	Biomaterials Biomedical Engineering Bioengineering Agricultural and Biological Sciences (miscellaneous) Biotechnology Research Article
Zdroj:	Synth Biol (Oxf)
ISSN:	2397-7000
Popis:	DNA templates for protein production remain an unexplored source of variability in the performance of cell-free expression (CFE) systems. 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 assess the concentration of the DNA template, the quality of the DNA template in terms of physical damage and the quality of the DNA solution in terms of purity resulting from eight DNA preparation workflows. We measure the variance in protein titer and rate of protein production in CFE reactions associated with the biological replicate of the DNA template, the technical replicate DNA solution prepared with the same workflow and the measurement replicate of nominally identical CFE reactions. We offer practical guidance for preparing and characterizing DNA templates to achieve acceptable variability in CFE performance.
Databáze:	OpenAIRE
Externí odkaz:	https://explore.openaire.eu/search/publication?articleId=doi_dedup___::8d9aed443cd1ab49ea60788e4e1b7f3f https://pubmed.ncbi.nlm.nih.gov/36046152 Zobrazit plný text záznamu Plný text ve formátu PDF Plný text ve formátu HTML
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