Cell-free translation: Preparation and validation of translation-competent extracts from saccharomyces cerevisiae

Brandon M. Trainor, Anton A. Komar, Dimitri G. Pestov, Natalia Shcherbik

Research output: Contribution to journalArticlepeer-review

3 Scopus citations

Abstract

Cell-free translation is a powerful technique for in vitro protein synthesis. While cell-free translation platforms prepared from bacterial, plant, and mammalian cells are commercially available, yeast-based translation systems remain proprietary knowledge of individual labs. Here, we provide a detailed protocol for simple, fast, and cost-effective preparation of the translation-competent cell-free extract (CFE) from budding yeast. Our protocol streamlines steps combined from different procedures published over the last three decades and incorporates cryogenic lysis of yeast cells to produce a high yield of the translationally active material. We also describe techniques for the validation and troubleshooting of the quality and translational activity of the obtained yeast CFE.

Original languageEnglish (US)
Article number4093
JournalBio-protocol
Volume11
Issue number18
DOIs
StatePublished - Sep 20 2021

All Science Journal Classification (ASJC) codes

  • General Biochemistry, Genetics and Molecular Biology
  • General Immunology and Microbiology
  • General Neuroscience
  • Plant Science

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