All chemicals used were reagent grade and purchased from Sigma-Aldrich (St. Louis, MO), unless otherwise noted. [EMIM]OAc, also known as [C2C1im]OAc, was purchased from Sigma-Aldrich and used as received. All strains and plasmids used in this work are listed at http://public-registry.jbei.org and are summarized in Additional file 5 : Table S4. E. coli DH10B (Invitrogen, Carlsbad, CA) was used for cloning and manipulation of plasmids. Except for the experiments described in Table 1 from the Keio Collection, all IL tolerance assays were conducted with E. coli DH1. Mutants from the Keio collection were tested against the BW25113 wild-type strain. Plasmids were constructed using isothermal assembly with 40 bp overlapping homologous sequences and plasmid sequences were verified with restriction endonuclease digestion and Sanger Sequencing (Quintara Biosciences, Albany, CA). We defined the promoter sequence of the cydDC operon as the immediate upstream ~ 500 bp of cydC. Plasmid transformation was conducted as previously described [60 (link)] or else as expressly indicated.
C2c1im oac
Manufactured by Merck Group
[C2C1im]OAc is an ionic liquid that serves as a versatile and efficient solvent for a variety of applications in the laboratory setting. As a salt-like compound, it exhibits unique properties that make it suitable for use in various chemical processes and analyses.
Automatically generated - may contain errors
Lab products found in correlation
E coli dh10b, by Thermo Fisher Scientific (1 mentions)
Emim oac, by Merck Group (1 mentions)
Crystal violet, by Thermo Fisher Scientific (1 mentions)
Zeocin, by Thermo Fisher Scientific (1 mentions)
Hygromycin b, by Thermo Fisher Scientific (1 mentions)
Geneticin, by Thermo Fisher Scientific (1 mentions)
C4c1im cl, by Merck Group (1 mentions)
C2c1im cl, by Merck Group (1 mentions)
Carbenicillin, by Chem-Impex (1 mentions)
2 protocols using c2c1im oac
1
Ionic Liquid Tolerance Engineering
2
Standard Yeast Media Preparation
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Standard yeast laboratory media were prepared as described elsewhere (Sherman 2002 (link)), with modifications. YPD (10 g/L yeast extract, 20 g/L peptone, 20 g/L dextrose) and synthetic complete (SC) media were adjusted to pH 5.0 with HCl. For experiments described in Supplemental Material, Figure S4, the pH was adjusted to the indicated values with HCl or NaOH accordingly. Cationic compounds were purchased from the following vendors: [C2C1im]Cl (catalog #272841, Sigma Aldrich; or catalog #AC354080250, Fisher Scientific, Pittsburgh, PA), [C4C1im]Cl (catalog #94128; Sigma Aldrich), [C2C1im][OAc] (catalog #689483; Sigma Aldrich), and Crystal Violet (CV) (catalog #NC9002731; Fisher Scientific). Cationic compounds were added directly to YPD or SC media and sterilized by passing through 2 μm filters. The following concentrations were used to select for plasmid and PCR product transformations in yeast and E. coli: 200 μg/ml Geneticin (catalog #10131027; Life Technologies), 100 μg/ml nourseothricin sulfate (catalog #RC-187; G-Biosciences), 200 μg/ml hygromycin B (catalog #10687010; GIBCO, Grand Island, NY), 200 μg/ml Zeocin (catalog #R25001; GIBCO), and 100 μg/ml carbenicillin (catalog #00049; Chem-Impex).
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