vitro protein synthesis  (New England Biolabs)

Article Title: Leveraging lipid-protein interactions to engineer spatial organization in cell-free systems
Article Snippet: .. Analysis of folding and insertion of cell-free expressed proteins into synthetic membranes: Protein expression was performed with the PURExpress In Vitro Protein Synthesis kit (E6800, NEB) according to the manufacturer's instructions. ..

Article Title: Localized ribosome access and distal tuning via the Listeria prfA RNA thermometer
Article Snippet: .. 350 ng of plasmid containing either prfA mutants or wild-type (WT), H4, or L5 were mixed with a final 1X PURExpress® solution from the PURExpress® In Vitro Protein Synthesis Kit (NEB #E6800), 0.7 μL RNaseOutTM Recombinant Ribonuclease Inhibitor (40 U/μL, Invitrogen #10777019), and 0.7 μL fluorescent lysine-charged tRNA from the FluoroTectTM GreenLys Translation Labeling System (Promega #L500A) for in-gel fluorescent visualization of protein expression. ..

Article Title: Similarity of the ion occupations of K + and Tl + in the selectivity filter of a K + channel, as verified by single-channel currents
Article Snippet: As described previously, the Kcv NTS gene [ , ] was cloned into a pEXP5-CT/TOPO®-vector (Invitrogen), and the fusion of the plasmid-encoded His tag was prevented by inserting a stop codon. .. In vitro expression was carried out using the PURExpress® In Vitro Protein Synthesis Kit (NEB), with one modification to the manufacturer’s protocol: Nanolipoproteins (MSP1D1-His/DMPC, Cube Biotech) were added to the expression mixture at a final concentration of 30 μM to enable proper folding of the hydrophobic Kcv NTS protein [ ]. .. Purification was performed using the His tag on the MSP1D1 scaffold protein with a 0.2-mL HisPur TM Ni-NTA spin column (ThermoFisher).

Article Title: Leveraging lipid-protein interactions to engineer spatial organization in cell-free systems
Article Snippet: .. Analysis of folding and insertion of cell-free expressed proteins into synthetic membranes: Protein expression was performed with the PURExpress In Vitro Protein Synthesis kit (E6800, NEB) according to the manufacturer's instructions. ..

Article Title: Coupling high-throughput protease enzymology with viral replication reveals biochemical constraints of viral fitness
Article Snippet: .. Protease–eGFP fusions were expressed in vitro using the PURExpress In Vitro Protein Synthesis Kit (NEB). .. Reactions (25 μL total) contained 10 μL Solution A, 7.5 μL Solution B, RNasin Ribonuclease Inhibitor (Promega), and 50-250 ng plasmid DNA.

Article Title: Development of a Microdroplet-Based Functional Genomic Screening pipeline by combination of DNA Nanoflowers and PURExpress Cell-Free Expression
Article Snippet: As a carrier-oil phase, HFE7500 (3MTM NovecTM 7500 Engineered Fluid) supplemented with 3% 008-fluorosurfactant from RAN biotechnologies, was used. .. As IVTT reconstituted system the PURExpress ® In Vitro Protein Synthesis Kit from NEB (MA, USA) was used following the manufacturer’s instructions and functioned as the aqueous phase during microdroplet generation. ..

Article Title: Evolution of Translation Initiation Factor 2 Extensions Links Initiation to Bacterial Stress Response
Article Snippet: .. For luminescence measurements, customized PURExpress In Vitro Protein Synthesis Kit (NEB) lacking endogenous IF2 was used (PURExpress-ΔIF2). ..

Article Title: 16S ribosomal RNA modification drives transcript-specific translation efficiency
Article Snippet: .. In vitro transcription/translation was performed using the PURExpress In Vitro Protein Synthesis kit (NEB) using ribosomes purified from WT or ΔmraW cells. ..

Article Title: Programmable Genome Sensing with Visual, Fluorescence, and Glucometer Readings: Cell-Free In Vitro Translation Approach Using RNA Switch-CRISPR
Article Snippet: All DNA and crRNA constructs were obtained from Integrated DNA Technologies, Inc. (Coralville, IA, USA) (see Table S1). .. PURExpress In Vitro Protein Synthesis Kit (catalog #E6800L), Monarch PCR & DNA Cleanup Kit (5 μg; catalog #T1030S), EnGen Lba Cas12a (Cas12a) (catalog #M0653), and NEBuffer 2.1 10X reaction buffer (catalog #B6002S) were purchased from New England Biolabs (Ipswich, MA, USA). .. The RPA kit (catalog #TABAS03KIT) was obtained from TwistDx Limited (Cambridge, UK).

Article Title: Localized ribosome access and distal tuning via the Listeria prfA RNA thermometer
Article Snippet: .. 350 ng of plasmid containing either prfA mutants or wild-type (WT), H4, or L5 were mixed with a final 1X PURExpress® solution from the PURExpress® In Vitro Protein Synthesis Kit (NEB #E6800), 0.7 μL RNaseOutTM Recombinant Ribonuclease Inhibitor (40 U/μL, Invitrogen #10777019), and 0.7 μL fluorescent lysine-charged tRNA from the FluoroTectTM GreenLys Translation Labeling System (Promega #L500A) for in-gel fluorescent visualization of protein expression. ..

Article Title: Similarity of the ion occupations of K + and Tl + in the selectivity filter of a K + channel, as verified by single-channel currents
Article Snippet: As described previously, the Kcv NTS gene [ , ] was cloned into a pEXP5-CT/TOPO®-vector (Invitrogen), and the fusion of the plasmid-encoded His tag was prevented by inserting a stop codon. .. In vitro expression was carried out using the PURExpress® In Vitro Protein Synthesis Kit (NEB), with one modification to the manufacturer’s protocol: Nanolipoproteins (MSP1D1-His/DMPC, Cube Biotech) were added to the expression mixture at a final concentration of 30 μM to enable proper folding of the hydrophobic Kcv NTS protein [ ]. .. Purification was performed using the His tag on the MSP1D1 scaffold protein with a 0.2-mL HisPur TM Ni-NTA spin column (ThermoFisher).

Article Title: 16S ribosomal RNA modification drives transcript-specific translation efficiency
Article Snippet: .. In vitro transcription/translation was performed using the PURExpress In Vitro Protein Synthesis kit (NEB) using ribosomes purified from WT or ΔmraW cells. ..

Article Title: Programmable Genome Sensing with Visual, Fluorescence, and Glucometer Readings: Cell-Free In Vitro Translation Approach Using RNA Switch-CRISPR
Article Snippet: All DNA and crRNA constructs were obtained from Integrated DNA Technologies, Inc. (Coralville, IA, USA) (see Table S1). .. PURExpress In Vitro Protein Synthesis Kit (catalog #E6800L), Monarch PCR & DNA Cleanup Kit (5 μg; catalog #T1030S), EnGen Lba Cas12a (Cas12a) (catalog #M0653), and NEBuffer 2.1 10X reaction buffer (catalog #B6002S) were purchased from New England Biolabs (Ipswich, MA, USA). .. The RPA kit (catalog #TABAS03KIT) was obtained from TwistDx Limited (Cambridge, UK).

Article Title: Localized ribosome access and distal tuning via the Listeria prfA RNA thermometer
Article Snippet: .. 350 ng of plasmid containing either prfA mutants or wild-type (WT), H4, or L5 were mixed with a final 1X PURExpress® solution from the PURExpress® In Vitro Protein Synthesis Kit (NEB #E6800), 0.7 μL RNaseOutTM Recombinant Ribonuclease Inhibitor (40 U/μL, Invitrogen #10777019), and 0.7 μL fluorescent lysine-charged tRNA from the FluoroTectTM GreenLys Translation Labeling System (Promega #L500A) for in-gel fluorescent visualization of protein expression. ..

Article Title: Localized ribosome access and distal tuning via the Listeria prfA RNA thermometer
Article Snippet: .. 350 ng of plasmid containing either prfA mutants or wild-type (WT), H4, or L5 were mixed with a final 1X PURExpress® solution from the PURExpress® In Vitro Protein Synthesis Kit (NEB #E6800), 0.7 μL RNaseOutTM Recombinant Ribonuclease Inhibitor (40 U/μL, Invitrogen #10777019), and 0.7 μL fluorescent lysine-charged tRNA from the FluoroTectTM GreenLys Translation Labeling System (Promega #L500A) for in-gel fluorescent visualization of protein expression. ..

Article Title: Localized ribosome access and distal tuning via the Listeria prfA RNA thermometer
Article Snippet: .. 350 ng of plasmid containing either prfA mutants or wild-type (WT), H4, or L5 were mixed with a final 1X PURExpress® solution from the PURExpress® In Vitro Protein Synthesis Kit (NEB #E6800), 0.7 μL RNaseOutTM Recombinant Ribonuclease Inhibitor (40 U/μL, Invitrogen #10777019), and 0.7 μL fluorescent lysine-charged tRNA from the FluoroTectTM GreenLys Translation Labeling System (Promega #L500A) for in-gel fluorescent visualization of protein expression. ..

Article Title: Similarity of the ion occupations of K + and Tl + in the selectivity filter of a K + channel, as verified by single-channel currents
Article Snippet: As described previously, the Kcv NTS gene [ , ] was cloned into a pEXP5-CT/TOPO®-vector (Invitrogen), and the fusion of the plasmid-encoded His tag was prevented by inserting a stop codon. .. In vitro expression was carried out using the PURExpress® In Vitro Protein Synthesis Kit (NEB), with one modification to the manufacturer’s protocol: Nanolipoproteins (MSP1D1-His/DMPC, Cube Biotech) were added to the expression mixture at a final concentration of 30 μM to enable proper folding of the hydrophobic Kcv NTS protein [ ]. .. Purification was performed using the His tag on the MSP1D1 scaffold protein with a 0.2-mL HisPur TM Ni-NTA spin column (ThermoFisher).

Article Title: Similarity of the ion occupations of K + and Tl + in the selectivity filter of a K + channel, as verified by single-channel currents
Article Snippet: As described previously, the Kcv NTS gene [ , ] was cloned into a pEXP5-CT/TOPO®-vector (Invitrogen), and the fusion of the plasmid-encoded His tag was prevented by inserting a stop codon. .. In vitro expression was carried out using the PURExpress® In Vitro Protein Synthesis Kit (NEB), with one modification to the manufacturer’s protocol: Nanolipoproteins (MSP1D1-His/DMPC, Cube Biotech) were added to the expression mixture at a final concentration of 30 μM to enable proper folding of the hydrophobic Kcv NTS protein [ ]. .. Purification was performed using the His tag on the MSP1D1 scaffold protein with a 0.2-mL HisPur TM Ni-NTA spin column (ThermoFisher).