TY  - JOUR
TI  - Cell-free Translation: Preparation and Validation of Translation-competent Extracts from Saccharomyces cerevisiae
AU  - Trainor, Brandon M.
AU  - Komar, Anton A.
AU  - Pestov, Dimitri G.
AU  - Shcherbik, Natalia 
VL  - 11
IS  - 18
PY  - 2021
DA  - 2021/09/20
SP  - e4093
C1  - Bio-protocol 2021;11:e4093
DO  - 10.21769/BioProtoc.4093
UR  - https://doi.org/10.21769/BioProtoc.4093
AB  - 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.Graphic abstract:The flow of Cell-Free Extract (CFE) preparation procedure.
KW  - Saccharomyces cerevisiae
KW  - Cell-free translation
KW  - Protein expression
KW  - Cryogenic lysis
KW  - Gel-filtration
KW  - RNA
JF  - Bio-protocol
SN  - 2331-8325
PB  - Bio-protocol LLC.
BIO101  - False