avian myelo blastosis virus reverse transcriptase  (New England Biolabs)

Article Title: The human branchpoint-interacting stem-loop sequence and structure regulates U2 snRNA expression, branchpoint recognition, and the transcriptome.
Article Snippet: A 5 ′ -end-labeled primer was generated by incubating 10 pmol of a DNA oligonucleotide complementary to nucleotides 39 –54 with γ- 32 P ATP and T4 Polynucleotide Kinase (Thermo Scientific cat. EK0031), followed by size exclusion with a Sephadex G-25 (Sigma –Aldrich, cat. G258050G) spin column. .. For primer extension reactions, 50 ng of U2 snRNA amplified cDNA and 0.1 picomole of labeled primer were incubated in 12 μl with annealing buffer (83.3 mM TrisHCl pH 7.9, 125 mM KCl) at 95 ◦C for 2 min, 37 ◦C (33 ◦C for P4 and P5 primers) for 10 min, and room temperature for 30 min, and then supplemented with 8 μl of extension mix to yield a final concentration of 10 mM DTT, 3 mM magnesium chloride, 0.1 mM dATP , dTTP , dCTP , and ddGTP and 2 units of AMV reverse transcriptase (NEB, cat. M0277S). ..

Article Title: The human branchpoint-interacting stem-loop sequence and structure regulates U2 snRNA expression, branchpoint recognition, and the transcriptome
Article Snippet: A 5′-end-labeled primer was generated by incubating 10 pmol of a DNA oligonucleotide complementary to nucleotides 39–54 with γ- 32 P ATP and T4 Polynucleotide Kinase (Thermo Scientific cat. EK0031), followed by size exclusion with a Sephadex G-25 (Sigma–Aldrich, cat. G2580-50G) spin column. .. For primer extension reactions, 50 ng of U2 snRNA amplified cDNA and 0.1 picomole of labeled primer were incubated in 12 μl with annealing buffer (83.3 mM Tris-HCl pH 7.9, 125 mM KCl) at 95°C for 2 min, 37°C (33°C for P4 and P5 primers) for 10 min, and room temperature for 30 min, and then supplemented with 8 μl of extension mix to yield a final concentration of 10 mM DTT, 3 mM magnesium chloride, 0.1 mM dATP, dTTP, dCTP, and ddGTP and 2 units of AMV reverse transcriptase (NEB, cat. M0277S). ..

Article Title: The human branchpoint-interacting stem-loop sequence and structure regulates U2 snRNA expression, branchpoint recognition, and the transcriptome.
Article Snippet: A 5 ′ -end-labeled primer was generated by incubating 10 pmol of a DNA oligonucleotide complementary to nucleotides 39 –54 with γ- 32 P ATP and T4 Polynucleotide Kinase (Thermo Scientific cat. EK0031), followed by size exclusion with a Sephadex G-25 (Sigma –Aldrich, cat. G258050G) spin column. .. For primer extension reactions, 50 ng of U2 snRNA amplified cDNA and 0.1 picomole of labeled primer were incubated in 12 μl with annealing buffer (83.3 mM TrisHCl pH 7.9, 125 mM KCl) at 95 ◦C for 2 min, 37 ◦C (33 ◦C for P4 and P5 primers) for 10 min, and room temperature for 30 min, and then supplemented with 8 μl of extension mix to yield a final concentration of 10 mM DTT, 3 mM magnesium chloride, 0.1 mM dATP , dTTP , dCTP , and ddGTP and 2 units of AMV reverse transcriptase (NEB, cat. M0277S). ..

Article Title: The human branchpoint-interacting stem-loop sequence and structure regulates U2 snRNA expression, branchpoint recognition, and the transcriptome
Article Snippet: A 5′-end-labeled primer was generated by incubating 10 pmol of a DNA oligonucleotide complementary to nucleotides 39–54 with γ- 32 P ATP and T4 Polynucleotide Kinase (Thermo Scientific cat. EK0031), followed by size exclusion with a Sephadex G-25 (Sigma–Aldrich, cat. G2580-50G) spin column. .. For primer extension reactions, 50 ng of U2 snRNA amplified cDNA and 0.1 picomole of labeled primer were incubated in 12 μl with annealing buffer (83.3 mM Tris-HCl pH 7.9, 125 mM KCl) at 95°C for 2 min, 37°C (33°C for P4 and P5 primers) for 10 min, and room temperature for 30 min, and then supplemented with 8 μl of extension mix to yield a final concentration of 10 mM DTT, 3 mM magnesium chloride, 0.1 mM dATP, dTTP, dCTP, and ddGTP and 2 units of AMV reverse transcriptase (NEB, cat. M0277S). ..

Article Title: Molecular mechanisms of PDCD4-mediated modulation of translation initiation and termination
Article Snippet: .. 2 μl buffer, containing 2.5 mM of each dNTPs, 25 mM Tris-HCl pH 7.5, 50 mM KAc, 2 mM DDT, 40 mM MgCl 2 , 250 nM 5’-6-FAM labeled primer 5’-GCATGTGCAGAGGACAGG-3’ (Syntol, Moscow, Russian Federation), 0.625 U AMV reverse transcriptase (NEB, Ipswich, USA), were added to ribosomal complexes. ..

Article Title: The human branchpoint-interacting stem-loop sequence and structure regulates U2 snRNA expression, branchpoint recognition, and the transcriptome.
Article Snippet: A 5 ′ -end-labeled primer was generated by incubating 10 pmol of a DNA oligonucleotide complementary to nucleotides 39 –54 with γ- 32 P ATP and T4 Polynucleotide Kinase (Thermo Scientific cat. EK0031), followed by size exclusion with a Sephadex G-25 (Sigma –Aldrich, cat. G258050G) spin column. .. For primer extension reactions, 50 ng of U2 snRNA amplified cDNA and 0.1 picomole of labeled primer were incubated in 12 μl with annealing buffer (83.3 mM TrisHCl pH 7.9, 125 mM KCl) at 95 ◦C for 2 min, 37 ◦C (33 ◦C for P4 and P5 primers) for 10 min, and room temperature for 30 min, and then supplemented with 8 μl of extension mix to yield a final concentration of 10 mM DTT, 3 mM magnesium chloride, 0.1 mM dATP , dTTP , dCTP , and ddGTP and 2 units of AMV reverse transcriptase (NEB, cat. M0277S). ..

Article Title: TOR-dependent regulation of the yeast homolog of the juvenile Batten Disease-associated gene CLN3
Article Snippet: .. AMV reverse transcriptase buffer, 8 units of murine RNase inhibitor (New England Biolabs, M0314), and 1 unit of AMV reverse transcriptase (New England Biolabs, M0277) were added, then incubated at 42 ∘ C for 1 hour, followed by 85 ∘ C for 5 minutes. .. The DNA was purified using SPRIselect magnetic beads (60 μ L) (Beckman Coulter) per reaction according to the manufacturer’s protocol.

Article Title: The human branchpoint-interacting stem-loop sequence and structure regulates U2 snRNA expression, branchpoint recognition, and the transcriptome
Article Snippet: A 5′-end-labeled primer was generated by incubating 10 pmol of a DNA oligonucleotide complementary to nucleotides 39–54 with γ- 32 P ATP and T4 Polynucleotide Kinase (Thermo Scientific cat. EK0031), followed by size exclusion with a Sephadex G-25 (Sigma–Aldrich, cat. G2580-50G) spin column. .. For primer extension reactions, 50 ng of U2 snRNA amplified cDNA and 0.1 picomole of labeled primer were incubated in 12 μl with annealing buffer (83.3 mM Tris-HCl pH 7.9, 125 mM KCl) at 95°C for 2 min, 37°C (33°C for P4 and P5 primers) for 10 min, and room temperature for 30 min, and then supplemented with 8 μl of extension mix to yield a final concentration of 10 mM DTT, 3 mM magnesium chloride, 0.1 mM dATP, dTTP, dCTP, and ddGTP and 2 units of AMV reverse transcriptase (NEB, cat. M0277S). ..

Article Title: Pseudouridylation landscape across 42 S. cerevisiae cytosolic tRNA isoacceptors via Nanopore direct RNA sequencing
Article Snippet: Primer extension reactions ( ) were done by annealing 1.25 μg total tRNA in 4 μL water (or 4 μL water alone for the no RNA control) with 1 μL of 1 μM IR800-labeled primer (IDT; Supplemental Table S6) at 95°C for 5 minutes on a heat block, then the samples (still in the block) were moved to room temperature to cool for 1 hour. .. Extension reactions were done by mixing 5 μL annealing reaction with 1 μL 10X AMV-RT buffer (NEB), 0.8 μL 5 mM dNTPs (NEB), 0.4 μL (4 units) AMV-Reverse Transcriptase (NEB), and 2.8 μL water, incubated at room temperature for 5 minutes, then at 37°C for two hours. .. Reactions were quenched by adding 10 μL 2X RNA dye (NEB) and stored at −20°C.

Article Title: The human branchpoint-interacting stem-loop sequence and structure regulates U2 snRNA expression, branchpoint recognition, and the transcriptome.
Article Snippet: A 5 ′ -end-labeled primer was generated by incubating 10 pmol of a DNA oligonucleotide complementary to nucleotides 39 –54 with γ- 32 P ATP and T4 Polynucleotide Kinase (Thermo Scientific cat. EK0031), followed by size exclusion with a Sephadex G-25 (Sigma –Aldrich, cat. G258050G) spin column. .. For primer extension reactions, 50 ng of U2 snRNA amplified cDNA and 0.1 picomole of labeled primer were incubated in 12 μl with annealing buffer (83.3 mM TrisHCl pH 7.9, 125 mM KCl) at 95 ◦C for 2 min, 37 ◦C (33 ◦C for P4 and P5 primers) for 10 min, and room temperature for 30 min, and then supplemented with 8 μl of extension mix to yield a final concentration of 10 mM DTT, 3 mM magnesium chloride, 0.1 mM dATP , dTTP , dCTP , and ddGTP and 2 units of AMV reverse transcriptase (NEB, cat. M0277S). ..

Article Title: The human branchpoint-interacting stem-loop sequence and structure regulates U2 snRNA expression, branchpoint recognition, and the transcriptome
Article Snippet: A 5′-end-labeled primer was generated by incubating 10 pmol of a DNA oligonucleotide complementary to nucleotides 39–54 with γ- 32 P ATP and T4 Polynucleotide Kinase (Thermo Scientific cat. EK0031), followed by size exclusion with a Sephadex G-25 (Sigma–Aldrich, cat. G2580-50G) spin column. .. For primer extension reactions, 50 ng of U2 snRNA amplified cDNA and 0.1 picomole of labeled primer were incubated in 12 μl with annealing buffer (83.3 mM Tris-HCl pH 7.9, 125 mM KCl) at 95°C for 2 min, 37°C (33°C for P4 and P5 primers) for 10 min, and room temperature for 30 min, and then supplemented with 8 μl of extension mix to yield a final concentration of 10 mM DTT, 3 mM magnesium chloride, 0.1 mM dATP, dTTP, dCTP, and ddGTP and 2 units of AMV reverse transcriptase (NEB, cat. M0277S). ..

Article Title: The human branchpoint-interacting stem-loop sequence and structure regulates U2 snRNA expression, branchpoint recognition, and the transcriptome.
Article Snippet: A 5 ′ -end-labeled primer was generated by incubating 10 pmol of a DNA oligonucleotide complementary to nucleotides 39 –54 with γ- 32 P ATP and T4 Polynucleotide Kinase (Thermo Scientific cat. EK0031), followed by size exclusion with a Sephadex G-25 (Sigma –Aldrich, cat. G258050G) spin column. .. For primer extension reactions, 50 ng of U2 snRNA amplified cDNA and 0.1 picomole of labeled primer were incubated in 12 μl with annealing buffer (83.3 mM TrisHCl pH 7.9, 125 mM KCl) at 95 ◦C for 2 min, 37 ◦C (33 ◦C for P4 and P5 primers) for 10 min, and room temperature for 30 min, and then supplemented with 8 μl of extension mix to yield a final concentration of 10 mM DTT, 3 mM magnesium chloride, 0.1 mM dATP , dTTP , dCTP , and ddGTP and 2 units of AMV reverse transcriptase (NEB, cat. M0277S). ..

Article Title: TOR-dependent regulation of the yeast homolog of the juvenile Batten Disease-associated gene CLN3
Article Snippet: .. AMV reverse transcriptase buffer, 8 units of murine RNase inhibitor (New England Biolabs, M0314), and 1 unit of AMV reverse transcriptase (New England Biolabs, M0277) were added, then incubated at 42 ∘ C for 1 hour, followed by 85 ∘ C for 5 minutes. .. The DNA was purified using SPRIselect magnetic beads (60 μ L) (Beckman Coulter) per reaction according to the manufacturer’s protocol.

Article Title: TOR-dependent regulation of the yeast homolog of the juvenile Batten Disease-associated gene CLN3
Article Snippet: .. The solution (10 μ L) was added to reaction buffer (10 μ L), AMV reverse transcriptase (10 U) (New England Biolabs, M0277), and murine RNase inhibitor, 8 U (New England Biolabs, M0314). ..

Article Title: The human branchpoint-interacting stem-loop sequence and structure regulates U2 snRNA expression, branchpoint recognition, and the transcriptome
Article Snippet: A 5′-end-labeled primer was generated by incubating 10 pmol of a DNA oligonucleotide complementary to nucleotides 39–54 with γ- 32 P ATP and T4 Polynucleotide Kinase (Thermo Scientific cat. EK0031), followed by size exclusion with a Sephadex G-25 (Sigma–Aldrich, cat. G2580-50G) spin column. .. For primer extension reactions, 50 ng of U2 snRNA amplified cDNA and 0.1 picomole of labeled primer were incubated in 12 μl with annealing buffer (83.3 mM Tris-HCl pH 7.9, 125 mM KCl) at 95°C for 2 min, 37°C (33°C for P4 and P5 primers) for 10 min, and room temperature for 30 min, and then supplemented with 8 μl of extension mix to yield a final concentration of 10 mM DTT, 3 mM magnesium chloride, 0.1 mM dATP, dTTP, dCTP, and ddGTP and 2 units of AMV reverse transcriptase (NEB, cat. M0277S). ..

Article Title: TOR-dependent regulation of the yeast homolog of the juvenile Batten Disease-associated gene CLN3
Article Snippet: We used poly A RNA and treated with DNase (New England Biolabs, M0303S) treated total RNA. .. AMV reverse transcriptase (New England Biolabs, M0277L) and RNase H (New England Biolabs M0297) were used to make the cDNA. .. The 5’ RACE PCR products from method A were PCR purified and tagged with adapter sequence, normalized and sent to Novogene, USA for NGS on NovaSeq X plus.

Article Title: Molecular mechanisms of PDCD4-mediated modulation of translation initiation and termination
Article Snippet: .. 2 μl buffer, containing 2.5 mM of each dNTPs, 25 mM Tris-HCl pH 7.5, 50 mM KAc, 2 mM DDT, 40 mM MgCl 2 , 250 nM 5’-6-FAM labeled primer 5’-GCATGTGCAGAGGACAGG-3’ (Syntol, Moscow, Russian Federation), 0.625 U AMV reverse transcriptase (NEB, Ipswich, USA), were added to ribosomal complexes. ..

Article Title: Infection of 5xFAD mice with a mouse‐adapted SARS‐CoV‐2 does not alter Alzheimer's disease neuropathology yet induces widespread changes in gene expression across diverse cell types
Article Snippet: .. cDNA was made following the “First Strand cDNA Synthesis” protocol by New England Biolabs, using AMV Reverse Transcriptase (New England Biolabs, M0277L), Random Hexamers (Invitrogen, N8080127), RNAse inhibitor (New England BiolabsM0314L), and AMV Buffer (New England Biolabs B0277A). qPCRs were performed using the QIAquant 96 2 plex and iTaq Universal SYBER Green Supermix (BioRad, 1725120). ..