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Promega tnt reaction buffer
Tnt Reaction Buffer, supplied by Promega, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/tnt+reaction+buffer/tnt+reaction+buffer/pmc05910820__pnas__1715888115__sapp-50-20-19
Average 90 stars, based on 1 article reviews
tnt reaction buffer - by Bioz Stars, 2026-09
90/100 stars

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Related Articles

Plasmid Preparation:

Article Title: DNA vaccination for treatment of multiple sclerosis and insulin-dependent diabetes mellitus
Article Snippet: .. Approximately 1 μg of plasmid DNA was incubated for 2 hours at 30° C. in a 50 μl volume containing the following: 25 μl of TNT rabbit reticulocyte lysate (Promega Corp., Madison, Wis.), 2 μl of TNT reaction buffer (Promega Corp., Madison, Wis.), 1 μl TNT T7 RNA polymerase (Promega Corp., Madison, Wis.), 1 μl of a 1 mM amino acid mixture minus methionine (Promega Corp., Madison, Wis.), 4 μl of 35S-methionine at 10 mCi/ml (Amersham Life Sciences Inc., Arlington Heights, Ill.), and 1 μl of RNasin ribonuclease inhibitor at 40 U/ml (Promega Corp., Madison, Wis.). ..

Article Title: Selective Removal of FG Repeat Domains from the Nuclear Pore Complex by Enterovirus 2A pro
Article Snippet: Point mutants were constructed at putative cleavage sites in Nup98 using the QuikChange II site-directed mutagenesis kit (Stratagene) and confirmed by sequencing. .. For in vitro translation, 0.5 μg of plasmid containing the wild-type (wt) or mutant Nup98 open reading frame was incubated with 1 μl of TnT reaction buffer (Promega), 12.5 μl of rabbit reticulocyte lysate, 0.5 μl of T7 RNA polymerase, 0.5 μl of 1 mM amino acid mixture without methionine, and 10 μCi of [ 35 S]methionine (>1,000 Ci/mmol; GE Healthcare) for 90 min at 30°C. ..

Article Title: Specific Cleavage of the Nuclear Pore Complex Protein Nup62 by a Viral Protease
Article Snippet: Mutagenesis and in Vitro Translation Point mutants were constructed at putative cleavage sites in Nup62 using the QuikChange II site-directed mutagenesis kit (Stratagene) and confirmed by sequencing. .. For in vitro translation, 0.5 μg of plasmid containing wild-type or mutant Nup62 open reading frame was incubated with 1 μl of TNT reaction buffer (Promega), 12.5 μl of rabbit reticulocyte lysate, 0.5 μl of T7 RNA polymerase, 0.5 μl of 1 m m amino acid mixture without methionine, and 10 μCi of [ 35 S]methionine (>1,000 Ci/mmol; GE Healthcare) for 90 min at 30 °C. ..

Article Title: Comprehensive, high-resolution binding energy landscapes reveal context dependencies of transcription factor binding
Article Snippet: 1.4 Protein expression using n vitro transcription and translation (IVTT) PHO4 and CBF1 open reading frames were sub-cloned into a pTNT vector (Promega/Genscript) with a C-terminal monomeric enhanced green fluorescent protein (meGFP) tag using Golden Gate Assembly, as previously reported (4, 5). .. Promega Wheat Germ Extract IVTT kit: • 50 μL of Promega TNT Wheat Germ Extract (L411A) • 4 μL Promega TNT Reaction Buffer (L462A) • 0.66 μL of Promega Amino Acid Mixture Minus Methionine, 1 mM (L996A) • 0.66 μL of Promega Amino Acid Mixture Minus Leucine, 1 mM (L995A) • 0.66 μL of Promega Amino Acid Mixture Minus Cysteine, 1 mM (L447C) • 2 μL of Promega Recombinant RNasin Ribonuclease Inhibitor (N251A) • 2 μL of Promega TNT T7 Wheat Germ Polymerase (L516A) • 1-2 μg of expression plasmid All components were allowed to equilibrate to 4◦C on ice. .. To a PCR strip tube, the following were added in order: 50 μL of Promega TNT Wheat Germ Extract, 4 μL of Promega TNT Reaction Buffer, 0.66 μL of 1 μM Promega Amino Acid Mixture Minus Methionine, 0.66 μL of 1 μM Promega Amino Acid Mixture Minus Leucine, 0.66 μL of 1 μM Promega Amino Acid Mixture Minus Cysteine, 2 μL of Promega Recombinant RNasin Ribonuclease Inhibitor, 2 μL of Promega TNT T7 Wheat Germ Polymerase, and 50 μL of DNase-/RNase-free water.

Article Title: Selective Removal of FG Repeat Domains from the Nuclear Pore Complex by Enterovirus 2A pro
Article Snippet: 173 Mutagenesis and in vitro translation- Point mutants were constructed at putative cleavage 174 sites in Nup98 using the QuickChange II Site-Directed Mutagenesis Kit (Stratagene) and 175 confirmed by sequencing. .. For in vitro translation, 0.5 μg of plasmid containing wild-type or 176 mutant Nup98 open reading frame was incubated with 1 μl of TnT reaction buffer (Promega), 177 12.5 μl of rabbit reticulocyte lysate, 0.5 μl of T7 RNA polymerase, 0.5 μl of 1 mM amino acid 178 mixture without methionine, and 10 μCi of 35S-methionine (>1,000 Ci/mmol, GE Healthcare) for 179 90 minutes at 30°C. ..

Article Title: Specific Cleavage of the Nuclear Pore Complex Protein Nup62 by a Viral Protease
Article Snippet: Mutagenesis and in Vitro Translation—Point mutants were constructed at putative cleavage sites in Nup62 using the QuikChange II site-directed mutagenesis kit (Stratagene) and confirmed by sequencing. .. For in vitro translation, 0.5 g of plasmid containing wild-type or mutant Nup62 open reading frame was incubated with 1 l of TNT reaction buffer (Promega), 12.5 l of rabbit reticulocyte lysate, 0.5 l of T7 RNA polymerase, 0.5 l of 1 mM amino acid mixture without methionine, and 10 Ci of [35S]methionine ( 1,000 Ci/mmol; GE Healthcare) for 90min at 30 °C. ..

Incubation:

Article Title: DNA vaccination for treatment of multiple sclerosis and insulin-dependent diabetes mellitus
Article Snippet: .. Approximately 1 μg of plasmid DNA was incubated for 2 hours at 30° C. in a 50 μl volume containing the following: 25 μl of TNT rabbit reticulocyte lysate (Promega Corp., Madison, Wis.), 2 μl of TNT reaction buffer (Promega Corp., Madison, Wis.), 1 μl TNT T7 RNA polymerase (Promega Corp., Madison, Wis.), 1 μl of a 1 mM amino acid mixture minus methionine (Promega Corp., Madison, Wis.), 4 μl of 35S-methionine at 10 mCi/ml (Amersham Life Sciences Inc., Arlington Heights, Ill.), and 1 μl of RNasin ribonuclease inhibitor at 40 U/ml (Promega Corp., Madison, Wis.). ..

Article Title: Selective Removal of FG Repeat Domains from the Nuclear Pore Complex by Enterovirus 2A pro
Article Snippet: Point mutants were constructed at putative cleavage sites in Nup98 using the QuikChange II site-directed mutagenesis kit (Stratagene) and confirmed by sequencing. .. For in vitro translation, 0.5 μg of plasmid containing the wild-type (wt) or mutant Nup98 open reading frame was incubated with 1 μl of TnT reaction buffer (Promega), 12.5 μl of rabbit reticulocyte lysate, 0.5 μl of T7 RNA polymerase, 0.5 μl of 1 mM amino acid mixture without methionine, and 10 μCi of [ 35 S]methionine (>1,000 Ci/mmol; GE Healthcare) for 90 min at 30°C. ..

Article Title: Specific Cleavage of the Nuclear Pore Complex Protein Nup62 by a Viral Protease
Article Snippet: Mutagenesis and in Vitro Translation Point mutants were constructed at putative cleavage sites in Nup62 using the QuikChange II site-directed mutagenesis kit (Stratagene) and confirmed by sequencing. .. For in vitro translation, 0.5 μg of plasmid containing wild-type or mutant Nup62 open reading frame was incubated with 1 μl of TNT reaction buffer (Promega), 12.5 μl of rabbit reticulocyte lysate, 0.5 μl of T7 RNA polymerase, 0.5 μl of 1 m m amino acid mixture without methionine, and 10 μCi of [ 35 S]methionine (>1,000 Ci/mmol; GE Healthcare) for 90 min at 30 °C. ..

Article Title: Selective Removal of FG Repeat Domains from the Nuclear Pore Complex by Enterovirus 2A pro
Article Snippet: 173 Mutagenesis and in vitro translation- Point mutants were constructed at putative cleavage 174 sites in Nup98 using the QuickChange II Site-Directed Mutagenesis Kit (Stratagene) and 175 confirmed by sequencing. .. For in vitro translation, 0.5 μg of plasmid containing wild-type or 176 mutant Nup98 open reading frame was incubated with 1 μl of TnT reaction buffer (Promega), 177 12.5 μl of rabbit reticulocyte lysate, 0.5 μl of T7 RNA polymerase, 0.5 μl of 1 mM amino acid 178 mixture without methionine, and 10 μCi of 35S-methionine (>1,000 Ci/mmol, GE Healthcare) for 179 90 minutes at 30°C. ..

Article Title: Specific Cleavage of the Nuclear Pore Complex Protein Nup62 by a Viral Protease
Article Snippet: Mutagenesis and in Vitro Translation—Point mutants were constructed at putative cleavage sites in Nup62 using the QuikChange II site-directed mutagenesis kit (Stratagene) and confirmed by sequencing. .. For in vitro translation, 0.5 g of plasmid containing wild-type or mutant Nup62 open reading frame was incubated with 1 l of TNT reaction buffer (Promega), 12.5 l of rabbit reticulocyte lysate, 0.5 l of T7 RNA polymerase, 0.5 l of 1 mM amino acid mixture without methionine, and 10 Ci of [35S]methionine ( 1,000 Ci/mmol; GE Healthcare) for 90min at 30 °C. ..

In Vitro:

Article Title: Selective Removal of FG Repeat Domains from the Nuclear Pore Complex by Enterovirus 2A pro
Article Snippet: Point mutants were constructed at putative cleavage sites in Nup98 using the QuikChange II site-directed mutagenesis kit (Stratagene) and confirmed by sequencing. .. For in vitro translation, 0.5 μg of plasmid containing the wild-type (wt) or mutant Nup98 open reading frame was incubated with 1 μl of TnT reaction buffer (Promega), 12.5 μl of rabbit reticulocyte lysate, 0.5 μl of T7 RNA polymerase, 0.5 μl of 1 mM amino acid mixture without methionine, and 10 μCi of [ 35 S]methionine (>1,000 Ci/mmol; GE Healthcare) for 90 min at 30°C. ..

Article Title: Specific Cleavage of the Nuclear Pore Complex Protein Nup62 by a Viral Protease
Article Snippet: Mutagenesis and in Vitro Translation Point mutants were constructed at putative cleavage sites in Nup62 using the QuikChange II site-directed mutagenesis kit (Stratagene) and confirmed by sequencing. .. For in vitro translation, 0.5 μg of plasmid containing wild-type or mutant Nup62 open reading frame was incubated with 1 μl of TNT reaction buffer (Promega), 12.5 μl of rabbit reticulocyte lysate, 0.5 μl of T7 RNA polymerase, 0.5 μl of 1 m m amino acid mixture without methionine, and 10 μCi of [ 35 S]methionine (>1,000 Ci/mmol; GE Healthcare) for 90 min at 30 °C. ..

Article Title: Selective Removal of FG Repeat Domains from the Nuclear Pore Complex by Enterovirus 2A pro
Article Snippet: 173 Mutagenesis and in vitro translation- Point mutants were constructed at putative cleavage 174 sites in Nup98 using the QuickChange II Site-Directed Mutagenesis Kit (Stratagene) and 175 confirmed by sequencing. .. For in vitro translation, 0.5 μg of plasmid containing wild-type or 176 mutant Nup98 open reading frame was incubated with 1 μl of TnT reaction buffer (Promega), 177 12.5 μl of rabbit reticulocyte lysate, 0.5 μl of T7 RNA polymerase, 0.5 μl of 1 mM amino acid 178 mixture without methionine, and 10 μCi of 35S-methionine (>1,000 Ci/mmol, GE Healthcare) for 179 90 minutes at 30°C. ..

Article Title: Specific Cleavage of the Nuclear Pore Complex Protein Nup62 by a Viral Protease
Article Snippet: Mutagenesis and in Vitro Translation—Point mutants were constructed at putative cleavage sites in Nup62 using the QuikChange II site-directed mutagenesis kit (Stratagene) and confirmed by sequencing. .. For in vitro translation, 0.5 g of plasmid containing wild-type or mutant Nup62 open reading frame was incubated with 1 l of TNT reaction buffer (Promega), 12.5 l of rabbit reticulocyte lysate, 0.5 l of T7 RNA polymerase, 0.5 l of 1 mM amino acid mixture without methionine, and 10 Ci of [35S]methionine ( 1,000 Ci/mmol; GE Healthcare) for 90min at 30 °C. ..

Mutagenesis:

Article Title: Selective Removal of FG Repeat Domains from the Nuclear Pore Complex by Enterovirus 2A pro
Article Snippet: Point mutants were constructed at putative cleavage sites in Nup98 using the QuikChange II site-directed mutagenesis kit (Stratagene) and confirmed by sequencing. .. For in vitro translation, 0.5 μg of plasmid containing the wild-type (wt) or mutant Nup98 open reading frame was incubated with 1 μl of TnT reaction buffer (Promega), 12.5 μl of rabbit reticulocyte lysate, 0.5 μl of T7 RNA polymerase, 0.5 μl of 1 mM amino acid mixture without methionine, and 10 μCi of [ 35 S]methionine (>1,000 Ci/mmol; GE Healthcare) for 90 min at 30°C. ..

Article Title: Specific Cleavage of the Nuclear Pore Complex Protein Nup62 by a Viral Protease
Article Snippet: Mutagenesis and in Vitro Translation Point mutants were constructed at putative cleavage sites in Nup62 using the QuikChange II site-directed mutagenesis kit (Stratagene) and confirmed by sequencing. .. For in vitro translation, 0.5 μg of plasmid containing wild-type or mutant Nup62 open reading frame was incubated with 1 μl of TNT reaction buffer (Promega), 12.5 μl of rabbit reticulocyte lysate, 0.5 μl of T7 RNA polymerase, 0.5 μl of 1 m m amino acid mixture without methionine, and 10 μCi of [ 35 S]methionine (>1,000 Ci/mmol; GE Healthcare) for 90 min at 30 °C. ..

Article Title: Selective Removal of FG Repeat Domains from the Nuclear Pore Complex by Enterovirus 2A pro
Article Snippet: 173 Mutagenesis and in vitro translation- Point mutants were constructed at putative cleavage 174 sites in Nup98 using the QuickChange II Site-Directed Mutagenesis Kit (Stratagene) and 175 confirmed by sequencing. .. For in vitro translation, 0.5 μg of plasmid containing wild-type or 176 mutant Nup98 open reading frame was incubated with 1 μl of TnT reaction buffer (Promega), 177 12.5 μl of rabbit reticulocyte lysate, 0.5 μl of T7 RNA polymerase, 0.5 μl of 1 mM amino acid 178 mixture without methionine, and 10 μCi of 35S-methionine (>1,000 Ci/mmol, GE Healthcare) for 179 90 minutes at 30°C. ..

Article Title: Specific Cleavage of the Nuclear Pore Complex Protein Nup62 by a Viral Protease
Article Snippet: Mutagenesis and in Vitro Translation—Point mutants were constructed at putative cleavage sites in Nup62 using the QuikChange II site-directed mutagenesis kit (Stratagene) and confirmed by sequencing. .. For in vitro translation, 0.5 g of plasmid containing wild-type or mutant Nup62 open reading frame was incubated with 1 l of TNT reaction buffer (Promega), 12.5 l of rabbit reticulocyte lysate, 0.5 l of T7 RNA polymerase, 0.5 l of 1 mM amino acid mixture without methionine, and 10 Ci of [35S]methionine ( 1,000 Ci/mmol; GE Healthcare) for 90min at 30 °C. ..

Recombinant:

Article Title: Comprehensive, high-resolution binding energy landscapes reveal context dependencies of transcription factor binding
Article Snippet: 1.4 Protein expression using n vitro transcription and translation (IVTT) PHO4 and CBF1 open reading frames were sub-cloned into a pTNT vector (Promega/Genscript) with a C-terminal monomeric enhanced green fluorescent protein (meGFP) tag using Golden Gate Assembly, as previously reported (4, 5). .. Promega Wheat Germ Extract IVTT kit: • 50 μL of Promega TNT Wheat Germ Extract (L411A) • 4 μL Promega TNT Reaction Buffer (L462A) • 0.66 μL of Promega Amino Acid Mixture Minus Methionine, 1 mM (L996A) • 0.66 μL of Promega Amino Acid Mixture Minus Leucine, 1 mM (L995A) • 0.66 μL of Promega Amino Acid Mixture Minus Cysteine, 1 mM (L447C) • 2 μL of Promega Recombinant RNasin Ribonuclease Inhibitor (N251A) • 2 μL of Promega TNT T7 Wheat Germ Polymerase (L516A) • 1-2 μg of expression plasmid All components were allowed to equilibrate to 4◦C on ice. .. To a PCR strip tube, the following were added in order: 50 μL of Promega TNT Wheat Germ Extract, 4 μL of Promega TNT Reaction Buffer, 0.66 μL of 1 μM Promega Amino Acid Mixture Minus Methionine, 0.66 μL of 1 μM Promega Amino Acid Mixture Minus Leucine, 0.66 μL of 1 μM Promega Amino Acid Mixture Minus Cysteine, 2 μL of Promega Recombinant RNasin Ribonuclease Inhibitor, 2 μL of Promega TNT T7 Wheat Germ Polymerase, and 50 μL of DNase-/RNase-free water.

Article Title: Comprehensive, high-resolution binding energy landscapes reveal context dependencies of transcription factor binding
Article Snippet: Promega Wheat Germ Extract IVTT kit: • 50 μL of Promega TNT Wheat Germ Extract (L411A) • 4 μL Promega TNT Reaction Buffer (L462A) • 0.66 μL of Promega Amino Acid Mixture Minus Methionine, 1 mM (L996A) • 0.66 μL of Promega Amino Acid Mixture Minus Leucine, 1 mM (L995A) • 0.66 μL of Promega Amino Acid Mixture Minus Cysteine, 1 mM (L447C) • 2 μL of Promega Recombinant RNasin Ribonuclease Inhibitor (N251A) • 2 μL of Promega TNT T7 Wheat Germ Polymerase (L516A) • 1-2 μg of expression plasmid All components were allowed to equilibrate to 4◦C on ice. .. To a PCR strip tube, the following were added in order: 50 μL of Promega TNT Wheat Germ Extract, 4 μL of Promega TNT Reaction Buffer, 0.66 μL of 1 μM Promega Amino Acid Mixture Minus Methionine, 0.66 μL of 1 μM Promega Amino Acid Mixture Minus Leucine, 0.66 μL of 1 μM Promega Amino Acid Mixture Minus Cysteine, 2 μL of Promega Recombinant RNasin Ribonuclease Inhibitor, 2 μL of Promega TNT T7 Wheat Germ Polymerase, and 50 μL of DNase-/RNase-free water. ..

Expressing:

Article Title: Comprehensive, high-resolution binding energy landscapes reveal context dependencies of transcription factor binding
Article Snippet: 1.4 Protein expression using n vitro transcription and translation (IVTT) PHO4 and CBF1 open reading frames were sub-cloned into a pTNT vector (Promega/Genscript) with a C-terminal monomeric enhanced green fluorescent protein (meGFP) tag using Golden Gate Assembly, as previously reported (4, 5). .. Promega Wheat Germ Extract IVTT kit: • 50 μL of Promega TNT Wheat Germ Extract (L411A) • 4 μL Promega TNT Reaction Buffer (L462A) • 0.66 μL of Promega Amino Acid Mixture Minus Methionine, 1 mM (L996A) • 0.66 μL of Promega Amino Acid Mixture Minus Leucine, 1 mM (L995A) • 0.66 μL of Promega Amino Acid Mixture Minus Cysteine, 1 mM (L447C) • 2 μL of Promega Recombinant RNasin Ribonuclease Inhibitor (N251A) • 2 μL of Promega TNT T7 Wheat Germ Polymerase (L516A) • 1-2 μg of expression plasmid All components were allowed to equilibrate to 4◦C on ice. .. To a PCR strip tube, the following were added in order: 50 μL of Promega TNT Wheat Germ Extract, 4 μL of Promega TNT Reaction Buffer, 0.66 μL of 1 μM Promega Amino Acid Mixture Minus Methionine, 0.66 μL of 1 μM Promega Amino Acid Mixture Minus Leucine, 0.66 μL of 1 μM Promega Amino Acid Mixture Minus Cysteine, 2 μL of Promega Recombinant RNasin Ribonuclease Inhibitor, 2 μL of Promega TNT T7 Wheat Germ Polymerase, and 50 μL of DNase-/RNase-free water.

Polymerase Chain Reaction:

Article Title: Comprehensive, high-resolution binding energy landscapes reveal context dependencies of transcription factor binding
Article Snippet: Promega Wheat Germ Extract IVTT kit: • 50 μL of Promega TNT Wheat Germ Extract (L411A) • 4 μL Promega TNT Reaction Buffer (L462A) • 0.66 μL of Promega Amino Acid Mixture Minus Methionine, 1 mM (L996A) • 0.66 μL of Promega Amino Acid Mixture Minus Leucine, 1 mM (L995A) • 0.66 μL of Promega Amino Acid Mixture Minus Cysteine, 1 mM (L447C) • 2 μL of Promega Recombinant RNasin Ribonuclease Inhibitor (N251A) • 2 μL of Promega TNT T7 Wheat Germ Polymerase (L516A) • 1-2 μg of expression plasmid All components were allowed to equilibrate to 4◦C on ice. .. To a PCR strip tube, the following were added in order: 50 μL of Promega TNT Wheat Germ Extract, 4 μL of Promega TNT Reaction Buffer, 0.66 μL of 1 μM Promega Amino Acid Mixture Minus Methionine, 0.66 μL of 1 μM Promega Amino Acid Mixture Minus Leucine, 0.66 μL of 1 μM Promega Amino Acid Mixture Minus Cysteine, 2 μL of Promega Recombinant RNasin Ribonuclease Inhibitor, 2 μL of Promega TNT T7 Wheat Germ Polymerase, and 50 μL of DNase-/RNase-free water. ..

Stripping Membranes:

Article Title: Comprehensive, high-resolution binding energy landscapes reveal context dependencies of transcription factor binding
Article Snippet: Promega Wheat Germ Extract IVTT kit: • 50 μL of Promega TNT Wheat Germ Extract (L411A) • 4 μL Promega TNT Reaction Buffer (L462A) • 0.66 μL of Promega Amino Acid Mixture Minus Methionine, 1 mM (L996A) • 0.66 μL of Promega Amino Acid Mixture Minus Leucine, 1 mM (L995A) • 0.66 μL of Promega Amino Acid Mixture Minus Cysteine, 1 mM (L447C) • 2 μL of Promega Recombinant RNasin Ribonuclease Inhibitor (N251A) • 2 μL of Promega TNT T7 Wheat Germ Polymerase (L516A) • 1-2 μg of expression plasmid All components were allowed to equilibrate to 4◦C on ice. .. To a PCR strip tube, the following were added in order: 50 μL of Promega TNT Wheat Germ Extract, 4 μL of Promega TNT Reaction Buffer, 0.66 μL of 1 μM Promega Amino Acid Mixture Minus Methionine, 0.66 μL of 1 μM Promega Amino Acid Mixture Minus Leucine, 0.66 μL of 1 μM Promega Amino Acid Mixture Minus Cysteine, 2 μL of Promega Recombinant RNasin Ribonuclease Inhibitor, 2 μL of Promega TNT T7 Wheat Germ Polymerase, and 50 μL of DNase-/RNase-free water. ..



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