Yeast tRNAPhe was purchased from Sigma-Aldrich. Other isoacceptor tRNAs were prepared from bulk tRNA (Roche) from either E. coli (tRNAVal, tRNALys, tRNAGln), and or S. cerevisiae (tRNAArg), via hybridization to immobilized complementary oligo DNAs as described previously49 (link),50 (link). E. coli and yeast tRNAs were charged with their cognate amino acids as described21 (link),27 (link),46 (link).
Yeast trnaphe
Manufactured by Merck Group
Sourced in United States
Yeast tRNAPhe is a type of transfer ribonucleic acid (tRNA) molecule isolated from the yeast Saccharomyces cerevisiae. It is responsible for the incorporation of the amino acid phenylalanine into the growing polypeptide chain during protein synthesis.
Automatically generated - may contain errors
Lab products found in correlation
Streptavidin c1 dynabeads, by Thermo Fisher Scientific (1 mentions)
Phosphoramidites, by Glen Research (1 mentions)
Rnase t1, by Worthington (1 mentions)
His tag protein purification kit, by Merck Group (1 mentions)
Dna oligonucleotide, by Integrated DNA Technologies (1 mentions)
Spermine s4264, by Merck Group (1 mentions)
Cis 4 hydroxy l proline, by Merck Group (1 mentions)
L azetidine 2 carboxylic acid, by Merck Group (1 mentions)
6 protocols using yeast trnaphe
1
Purification and Charging of tRNA Isoacceptors
2
In Vitro Transcription and Synthesis of Nucleic Acid Targets
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The polymerase ribozyme and all RNA templates were prepared by in vitro transcription, as described previously (14 (link)). Target DNAs were purchased from IDT. Most target RNAs were prepared by solid-phase synthesis employing phosphoramidites and other reagents from Glen Research (Sterling, VA, USA), with the exception of yeast tRNAPhe, which was purchased from Sigma-Aldrich (St. Louis, MO, USA). All modified NTPs and dNTPs were purchased from either Jena Bioscience (Jena, Germany) or TriLink BioTechnologies (San Diego, CA, USA). Streptavidin C1 Dynabeads were purchased from ThermoFisher (Grand Island, NY, USA).
3
Customized RNA Sequence Analysis
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A customized RNA (5′-GCAUCAGAAAUACACCCGUAGGGCUUU-GAGA-3′) with all the dinucleotide combinations (42 = 16) and four homopolymer sequences was purchased from Integrated DNA Technology (IDT). Yeast tRNAPhe was obtained from Sigma, and all other chemicals were procured from Thermo Fisher Scientific. RNase T1 was obtained from Worthington Biochemical Corporation. RNase MC1 [11 (link)] and cusativin [12 (link)] were purified on a nickel column using His-tag protein purification kit (EMD Millipore) following their expression from recombinant plasmids in Rosetta TM (DE3) and BL21 strains of Escherichia coli, as described before.
4
Proline Analogs and Polyamine Purification
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Proline and its analogs, α-methyl-l -proline, 3,4-dehydro-dl -proline, cis-4-hydroxy-l -proline and l -azetidine-2-carboxylic acid, as well as spermidine (catalog number S2626), spermine (S4264) and putrescine (P5780), and yeast tRNAPhe were purchased from Sigma. All DNA oligonucleotides were purchased from either Integrated DNA Technologies or Eurofins MWG Operon. The 3′-biotin-labeled RNA oligonucleotide used for affinity purification of yeast tRNAPro was also purchased from Eurofins MWG Operon; yeast tRNALys was obtained from tRNA Probes (College Station, TX, USA).
5
Purification and Preparation of RNA for Structural Studies
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Yeast tRNAPhe was purchased from Sigma-Aldrich (R4018). rU20 was purchased from Integrated DNA Technologies. T7-transcribed, stabilized P4–P6 RNA was produced as in Athavale et al. (47 (link)). Background Mg2+ was removed from the RNAs by dialysis in 180 mM NaCl and 50 mM HEPES buffer pH 7.1 using a 10 kDa MWCO filter.
6
Purification and Charging of Isoacceptor tRNAs
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Yeast tRNAPhe was purchased from Sigma-Aldrich. Other isoacceptor tRNAs were prepared from bulk tRNA (Roche) from either E. coli (tRNAVal, tRNALys, tRNAGln, and elongator tRNAMet) or yeast (tRNAArg, tRNATrp, and tRNALeu) via hybridization to immobilized complementary oligoDNAs as described previously (1 (link), 39 (link)). E. coli and yeast tRNAs were charged with their cognate amino acids as described (56 (link), 57 (link)). E. coli tRNAGln was charged with either [3H]-Gln for the peptide release assay or [14C]-Gln for the Trp-tRNATrp(Prf) binding assay. Yeast tRNATrp was charged with [3H]-Trp for the hexapeptide assay or the hexapeptidyl-tRNA dissociation assay. E. coli elongator tRNAMet was charged with [35S]-Met for the octapeptide formation assay.
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