title: Preparation of amino acid mixtures for cell-free expression systems.
abstract: Here we present a procedure for preparing amino acid mixtures--having both the desired composition and a physiological pH--at high concentrations for cell-free expression systems. Up to 2.1 mg/mL of active protein was synthesized in batch mode reactions with an all Escherichia coli cell-free expression system. Our method is fast, easy to execute, and economically advantageous compared to expensive commercial kits, making it useful for high-throughput experiments, incorporation of nonstandard amino acids, and cell-free metabolic engineering.
authors.affiliation: Department of Physics, University of Minnesota, Minneapolis, MN.
authors.forename: Filippo
authors.initials: F
authors.lastname: Caschera
authors.affiliation: Department of Physics, University of Minnesota, Minneapolis, MN.
authors.forename: Vincent
authors.initials: V
authors.lastname: Noireaux
mesh_terms: D000596:Amino Acids / Q000737:chemistry*; D002474:Cell-Free System; D004926:Escherichia coli / Q000378:metabolism; D060847:Metabolic Engineering / Q000379:methods*
chemical_list: D000596:Amino Acids
keywords: amino acid distribution; amino acid mixtures; cell-free transcription-translation; pH