Rosetta 2 cells containing pET22b-U2 were obtained in house.[8 (link)] Degenerate primers for site saturation mutagenesis of position 49 were purchased from Integrated DNA Technologies (Skokie, Illinois, USA). Dpn1 enzyme was purchased from New England Biolabs (Ipswich, Massachusetts, USA). Saccharomyces cerevisiae tRNAPhe, terrific broth media, hexafluoroisopropanol (HFIP), and ammonium acetate solution were obtained from Sigma Aldrich (St. Louis, Missouri, USA). Carbenicillin, chloramphenicol, Isopropyl-β-D-thiogalactoside (IPTG), and lysozyme were purchased from Gold Biotechnology (St. Louis, Missouri, USA). The Ni-NTA His-Bind purification kit and 4–20% SDS gels were purchased from Novagen (Madison, Wisconsin, USA). Hydrochloric acid, 15% TBE-urea gels, Tris-Cl, boric acid, and triethylamine (TEA) were acquired from Fisher Scientific (Waltham, Massachusetts, USA). The synthetic 12-mer oligonucleotide was purchased from Dharmacon (Lafayette, Colorado, USA).
Saccharomyces cerevisiae trnaphe
Manufactured by Merck Group
Sourced in United States
Saccharomyces cerevisiae tRNAPhe is a transfer RNA (tRNA) molecule isolated from the yeast Saccharomyces cerevisiae. It is responsible for the incorporation of the amino acid phenylalanine during protein synthesis.
Automatically generated - may contain errors
Lab products found in correlation
Ammonium acetate solution, by Merck Group (1 mentions)
Hydrochloric acid (hcl), by Thermo Fisher Scientific (1 mentions)
Carbenicillin, by GoldBio (1 mentions)
Boric acid, by Thermo Fisher Scientific (1 mentions)
Chloramphenicol, by GoldBio (1 mentions)
Lysozyme, by GoldBio (1 mentions)
Dpn1 enzyme, by New England Biolabs (1 mentions)
Isopropyl β d thiogalactoside, by GoldBio (1 mentions)
Tris cl, by Thermo Fisher Scientific (1 mentions)
Hexafluoroisopropanol hfip, by Merck Group (1 mentions)
Triethylamine tea, by Thermo Fisher Scientific (1 mentions)
2 protocols using saccharomyces cerevisiae trnaphe
1
Site-Directed Mutagenesis Protocol for Protein Engineering
2
Purification of E. coli GluRS and tRNA_Glu
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Overproduction and purification of E. coli GluRS were performed as previously described [19 (link)] with the following modifications. A C-terminal histidine-tagged GluRS was used instead of the N-terminal tagged one. The overproduction was induced overnight at 30°C with 1 mM IPTG. The GluRS was purified to homogeneity, as revealed by SDS-PAGE analysis (result not shown).
Overproduction and purification of E. coli tRNAGlu-enriched total tRNA was done as described [20 (link)]. The aminoacylation plateau indicated that the final product contained 26% tRNAGlu. Saccharomyces cerevisiae tRNAPhe, used as a negative control, was purchased from Sigma-Aldrich (cat No: R4018).
Overproduction and purification of E. coli tRNAGlu-enriched total tRNA was done as described [20 (link)]. The aminoacylation plateau indicated that the final product contained 26% tRNAGlu. Saccharomyces cerevisiae tRNAPhe, used as a negative control, was purchased from Sigma-Aldrich (cat No: R4018).
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