Structured Review

Promega t4 ligase buffer
T4 Ligase Buffer, supplied by Promega, used in various techniques. Bioz Stars score: 99/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/result/t4 ligase buffer/product/Promega
Average 99 stars, based on 3 article reviews
Price from $9.99 to $1999.99
t4 ligase buffer - by Bioz Stars, 2020-05
99/100 stars

Images

Related Articles

Amplification:

Article Title: Bovine Leukemia Virus Structural Gene Vectors Are Immunogenic and Lack Pathogenicity in a Rabbit Model
Article Snippet: .. One microgram of PBMC total RNA was subjected to reverse transcription for 1 h at 48°C and PCR amplification in the single-buffer Access reverse transcription-PCR (RT-PCR) system (Promega) with BLV pol primers KB2341 and KB3175. .. Nested PCR with 1/50 of the primary PCR product was performed with BLV pol primers KB560 and KB561.

Polymerase Chain Reaction:

Article Title: Bovine Leukemia Virus Structural Gene Vectors Are Immunogenic and Lack Pathogenicity in a Rabbit Model
Article Snippet: .. One microgram of PBMC total RNA was subjected to reverse transcription for 1 h at 48°C and PCR amplification in the single-buffer Access reverse transcription-PCR (RT-PCR) system (Promega) with BLV pol primers KB2341 and KB3175. .. Nested PCR with 1/50 of the primary PCR product was performed with BLV pol primers KB560 and KB561.

Article Title: Genomic organization and promoter analysis of the human heat shock factor 2 gene
Article Snippet: .. The PCR reaction was carried out under the following conditions: primers (300 nM); 0.2–0.3 μg genomic P1 plasmid; 1X Pfu-PCR buffer containing MgCl2 (1.5 mM, Promega); dATP, dGTP, dCTP, and dTTP (0.2 mM each); and Pfu DNA polymerase (1.25 U, Promega) in a volume of 50 μL. ..

Protease Inhibitor:

Article Title: The roles of RRP15 in nucleolar formation, ribosome biogenesis and checkpoint control in human cells
Article Snippet: .. The nuclear lysate was extracted in extraction buffer (25 mM Tris (pH 7.5), 100 mM KCl, 1 mM DTT, 2 mM EDTA, 0.1% NP-40, 1 mM NaF, 40 mg/ml of phenylmethylsulfonyl fluoride, 10 mg/ml of protease inhibitor cocktail and 0.1U/ml of RNasin (Promega)) and sonicated. .. The nuclear lysate was overlaid on 10 to 30% (wt/wt) sucrose gradients in preribosome buffer (25 mM Tris (pH 7.5), 100 mM KCl, 1 mM DTT and 2 mM EDTA) and centrifuged at 36,000 rpm for 3 h at 4°C in a Beckman SW41Ti rotor.

Reverse Transcription Polymerase Chain Reaction:

Article Title: Bovine Leukemia Virus Structural Gene Vectors Are Immunogenic and Lack Pathogenicity in a Rabbit Model
Article Snippet: .. One microgram of PBMC total RNA was subjected to reverse transcription for 1 h at 48°C and PCR amplification in the single-buffer Access reverse transcription-PCR (RT-PCR) system (Promega) with BLV pol primers KB2341 and KB3175. .. Nested PCR with 1/50 of the primary PCR product was performed with BLV pol primers KB560 and KB561.

Sequencing:

Article Title: Aspergillus niger membrane-associated proteome analysis for the identification of glucose transporters
Article Snippet: .. Approximately 150 μL of digestion buffer, containing sequencing grade modified trypsin (12.5 ng μL−1 ) (Promega, Madison, WI, USA) in ABC, was added to each sample, making sure that all gel pieces were kept wet during digestion (adding, if necessary, additional ABC solution). .. Peptide digestion products were extracted by adding 50 μL of 2 % trifluoroacetic acid (TFA), followed by an incubation step in a thermoblock (ThermoMixer) for 20 min, at room temperature and vigorous stirring (1400 rpm).

Isolation:

Article Title: Mapping targets for small nucleolar RNAs in yeast
Article Snippet: .. RNA-protein complexes were isolated by binding to an IgG column (GE), washed in buffer A and released by TEV (Promega) cleavage. .. RNAs were partially digested using RNaceIT Ribonuclease Cocktail (Agilent).

Modification:

Article Title: Aspergillus niger membrane-associated proteome analysis for the identification of glucose transporters
Article Snippet: .. Approximately 150 μL of digestion buffer, containing sequencing grade modified trypsin (12.5 ng μL−1 ) (Promega, Madison, WI, USA) in ABC, was added to each sample, making sure that all gel pieces were kept wet during digestion (adding, if necessary, additional ABC solution). .. Peptide digestion products were extracted by adding 50 μL of 2 % trifluoroacetic acid (TFA), followed by an incubation step in a thermoblock (ThermoMixer) for 20 min, at room temperature and vigorous stirring (1400 rpm).

Sonication:

Article Title: The roles of RRP15 in nucleolar formation, ribosome biogenesis and checkpoint control in human cells
Article Snippet: .. The nuclear lysate was extracted in extraction buffer (25 mM Tris (pH 7.5), 100 mM KCl, 1 mM DTT, 2 mM EDTA, 0.1% NP-40, 1 mM NaF, 40 mg/ml of phenylmethylsulfonyl fluoride, 10 mg/ml of protease inhibitor cocktail and 0.1U/ml of RNasin (Promega)) and sonicated. .. The nuclear lysate was overlaid on 10 to 30% (wt/wt) sucrose gradients in preribosome buffer (25 mM Tris (pH 7.5), 100 mM KCl, 1 mM DTT and 2 mM EDTA) and centrifuged at 36,000 rpm for 3 h at 4°C in a Beckman SW41Ti rotor.

Binding Assay:

Article Title: Mapping targets for small nucleolar RNAs in yeast
Article Snippet: .. RNA-protein complexes were isolated by binding to an IgG column (GE), washed in buffer A and released by TEV (Promega) cleavage. .. RNAs were partially digested using RNaceIT Ribonuclease Cocktail (Agilent).

SDS Page:

Article Title: A novel mechanism of RNase L inhibition: Theiler's virus L* protein prevents 2-5A from binding to RNase L
Article Snippet: .. The peak fractions containing L* were pooled and diluted 1:3 in buffer A and bound with Ni-NTA resin, washed with buffer A containing 10 mM imidazole and eluted with 150 mM imidazole in buffer A. Fractions were loaded on 12% SDS-PAGE and stained with Gel-Code Blue (Promega). .. Human RNase L was produced as described [ ].

Plasmid Preparation:

Article Title: Genomic organization and promoter analysis of the human heat shock factor 2 gene
Article Snippet: .. The PCR reaction was carried out under the following conditions: primers (300 nM); 0.2–0.3 μg genomic P1 plasmid; 1X Pfu-PCR buffer containing MgCl2 (1.5 mM, Promega); dATP, dGTP, dCTP, and dTTP (0.2 mM each); and Pfu DNA polymerase (1.25 U, Promega) in a volume of 50 μL. ..

Staining:

Article Title: A novel mechanism of RNase L inhibition: Theiler's virus L* protein prevents 2-5A from binding to RNase L
Article Snippet: .. The peak fractions containing L* were pooled and diluted 1:3 in buffer A and bound with Ni-NTA resin, washed with buffer A containing 10 mM imidazole and eluted with 150 mM imidazole in buffer A. Fractions were loaded on 12% SDS-PAGE and stained with Gel-Code Blue (Promega). .. Human RNase L was produced as described [ ].

Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 92
    Promega t4 dna ligase buffer
    DNA supercoiling and bending assays by phosphorylated SmHMGB1. (A) Circular relaxed plasmid pTZ19R DNA was incubated in the presence of topoisomerase I with 1 µg of recombinant SmHMGB1-FL or SmHMGB1-S172A/S174A that were phosphorylated (lanes 3–5) or not (lanes 6–8 and 9–11), by CK2. Deproteinized DNA topoisomers were resolved on 1% agarose gels, followed by staining of the gels with ethidium bromide. Form I, supercoiled DNA; form II, relaxed circular DNA. (B) Top panel: autoradiography; bottom panel: Coomassie staining. (C) A 32 P-labeled 123-bp DNA fragment (∼1 nM) was pre-incubated with 50 ng of recombinant proteins, that were phosphorylated (lanes 7–9) or not (lanes 4–6, 10–12, 13–15 and 16–18), followed by ligation with <t>T4</t> DNA ligase. Exonuclease III was used to verify the identity of DNA circles. The deproteinized DNA ligation products were subjected to electrophoresis on 6% non-denaturing polyacrylamide gels and visualized by autoradiography. Controls are as follows: FL(c1): SmHMGB1-FL without CK2; FL(c2): SmHMGB1-FL without phosphate; FL(c3): SmHMGB1-FL without CK2 buffer. Linear: linear DNA; Lm: linear multimers. (D) Top panel: autoradiography; bottom panel: Coomassie staining. These experiments were repeated four times.
    T4 Dna Ligase Buffer, supplied by Promega, used in various techniques. Bioz Stars score: 92/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/t4 dna ligase buffer/product/Promega
    Average 92 stars, based on 15 article reviews
    Price from $9.99 to $1999.99
    t4 dna ligase buffer - by Bioz Stars, 2020-05
    92/100 stars
      Buy from Supplier

    91
    Promega t4 ligase buffer
    DNA supercoiling and bending assays by phosphorylated SmHMGB1. (A) Circular relaxed plasmid pTZ19R DNA was incubated in the presence of topoisomerase I with 1 µg of recombinant SmHMGB1-FL or SmHMGB1-S172A/S174A that were phosphorylated (lanes 3–5) or not (lanes 6–8 and 9–11), by CK2. Deproteinized DNA topoisomers were resolved on 1% agarose gels, followed by staining of the gels with ethidium bromide. Form I, supercoiled DNA; form II, relaxed circular DNA. (B) Top panel: autoradiography; bottom panel: Coomassie staining. (C) A 32 P-labeled 123-bp DNA fragment (∼1 nM) was pre-incubated with 50 ng of recombinant proteins, that were phosphorylated (lanes 7–9) or not (lanes 4–6, 10–12, 13–15 and 16–18), followed by ligation with <t>T4</t> DNA ligase. Exonuclease III was used to verify the identity of DNA circles. The deproteinized DNA ligation products were subjected to electrophoresis on 6% non-denaturing polyacrylamide gels and visualized by autoradiography. Controls are as follows: FL(c1): SmHMGB1-FL without CK2; FL(c2): SmHMGB1-FL without phosphate; FL(c3): SmHMGB1-FL without CK2 buffer. Linear: linear DNA; Lm: linear multimers. (D) Top panel: autoradiography; bottom panel: Coomassie staining. These experiments were repeated four times.
    T4 Ligase Buffer, supplied by Promega, used in various techniques. Bioz Stars score: 91/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/t4 ligase buffer/product/Promega
    Average 91 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    t4 ligase buffer - by Bioz Stars, 2020-05
    91/100 stars
      Buy from Supplier

    93
    Promega t4 rna ligase
    DNA supercoiling and bending assays by phosphorylated SmHMGB1. (A) Circular relaxed plasmid pTZ19R DNA was incubated in the presence of topoisomerase I with 1 µg of recombinant SmHMGB1-FL or SmHMGB1-S172A/S174A that were phosphorylated (lanes 3–5) or not (lanes 6–8 and 9–11), by CK2. Deproteinized DNA topoisomers were resolved on 1% agarose gels, followed by staining of the gels with ethidium bromide. Form I, supercoiled DNA; form II, relaxed circular DNA. (B) Top panel: autoradiography; bottom panel: Coomassie staining. (C) A 32 P-labeled 123-bp DNA fragment (∼1 nM) was pre-incubated with 50 ng of recombinant proteins, that were phosphorylated (lanes 7–9) or not (lanes 4–6, 10–12, 13–15 and 16–18), followed by ligation with <t>T4</t> DNA ligase. Exonuclease III was used to verify the identity of DNA circles. The deproteinized DNA ligation products were subjected to electrophoresis on 6% non-denaturing polyacrylamide gels and visualized by autoradiography. Controls are as follows: FL(c1): SmHMGB1-FL without CK2; FL(c2): SmHMGB1-FL without phosphate; FL(c3): SmHMGB1-FL without CK2 buffer. Linear: linear DNA; Lm: linear multimers. (D) Top panel: autoradiography; bottom panel: Coomassie staining. These experiments were repeated four times.
    T4 Rna Ligase, supplied by Promega, used in various techniques. Bioz Stars score: 93/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/t4 rna ligase/product/Promega
    Average 93 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    t4 rna ligase - by Bioz Stars, 2020-05
    93/100 stars
      Buy from Supplier

    Image Search Results


    DNA supercoiling and bending assays by phosphorylated SmHMGB1. (A) Circular relaxed plasmid pTZ19R DNA was incubated in the presence of topoisomerase I with 1 µg of recombinant SmHMGB1-FL or SmHMGB1-S172A/S174A that were phosphorylated (lanes 3–5) or not (lanes 6–8 and 9–11), by CK2. Deproteinized DNA topoisomers were resolved on 1% agarose gels, followed by staining of the gels with ethidium bromide. Form I, supercoiled DNA; form II, relaxed circular DNA. (B) Top panel: autoradiography; bottom panel: Coomassie staining. (C) A 32 P-labeled 123-bp DNA fragment (∼1 nM) was pre-incubated with 50 ng of recombinant proteins, that were phosphorylated (lanes 7–9) or not (lanes 4–6, 10–12, 13–15 and 16–18), followed by ligation with T4 DNA ligase. Exonuclease III was used to verify the identity of DNA circles. The deproteinized DNA ligation products were subjected to electrophoresis on 6% non-denaturing polyacrylamide gels and visualized by autoradiography. Controls are as follows: FL(c1): SmHMGB1-FL without CK2; FL(c2): SmHMGB1-FL without phosphate; FL(c3): SmHMGB1-FL without CK2 buffer. Linear: linear DNA; Lm: linear multimers. (D) Top panel: autoradiography; bottom panel: Coomassie staining. These experiments were repeated four times.

    Journal: PLoS ONE

    Article Title: CK2 Phosphorylation of Schistosoma mansoni HMGB1 Protein Regulates Its Cellular Traffic and Secretion but Not Its DNA Transactions

    doi: 10.1371/journal.pone.0023572

    Figure Lengend Snippet: DNA supercoiling and bending assays by phosphorylated SmHMGB1. (A) Circular relaxed plasmid pTZ19R DNA was incubated in the presence of topoisomerase I with 1 µg of recombinant SmHMGB1-FL or SmHMGB1-S172A/S174A that were phosphorylated (lanes 3–5) or not (lanes 6–8 and 9–11), by CK2. Deproteinized DNA topoisomers were resolved on 1% agarose gels, followed by staining of the gels with ethidium bromide. Form I, supercoiled DNA; form II, relaxed circular DNA. (B) Top panel: autoradiography; bottom panel: Coomassie staining. (C) A 32 P-labeled 123-bp DNA fragment (∼1 nM) was pre-incubated with 50 ng of recombinant proteins, that were phosphorylated (lanes 7–9) or not (lanes 4–6, 10–12, 13–15 and 16–18), followed by ligation with T4 DNA ligase. Exonuclease III was used to verify the identity of DNA circles. The deproteinized DNA ligation products were subjected to electrophoresis on 6% non-denaturing polyacrylamide gels and visualized by autoradiography. Controls are as follows: FL(c1): SmHMGB1-FL without CK2; FL(c2): SmHMGB1-FL without phosphate; FL(c3): SmHMGB1-FL without CK2 buffer. Linear: linear DNA; Lm: linear multimers. (D) Top panel: autoradiography; bottom panel: Coomassie staining. These experiments were repeated four times.

    Article Snippet: Briefly, a 32 P-labeled-66-bp or a 32 P-labeled-123-bp DNA fragments (1 nM) with cohesive BamHI ends were pre-incubated on ice for 20 min with appropriate amounts of recombinant proteins (50 ng), total (10 µg), nuclear (4 µg) or cytoplasmic (4 µg) adult worm extracts, in 1× T4 DNA ligase buffer (30 mM Tris–HCl, pH 7.8, 10 mM MgCl2 , 10 mM dithiothreitol, and 0.5 mM ATP; Promega) in a final volume of 20 µl.

    Techniques: Plasmid Preparation, Incubation, Recombinant, Staining, Autoradiography, Labeling, Ligation, DNA Ligation, Electrophoresis

    DNA transactions by recombinant AaHMGB1 proteins. (A) Preferential binding of AaHMGB1 protein to supercoiled DNA. An equimolar mixture of supercoiled and linearized plasmid pTZ19R (∼10 nM) was pre-incubated with increasing amounts of AaHMGB1 (0.5–1 µM) and the DNA–protein complexes were resolved on a 1% agarose gel, followed by staining of the gel with ethidium bromide. Form I, supercoiled DNA; L, Linear DNA; Form II, relaxed circular DNA; (B) DNA supercoiling by AaHMGB1 and its truncated forms. Circular relaxed plasmid pTZ19R DNA was incubated in the presence of topoisomerase I (Topo I) and AaHMGB1 recombinant proteins (7–14 µM). Deproteinized DNA topoisomers were resolved on 1% agarose gels, followed by staining of the gel with ethidium bromide. Form I, supercoiled DNA; Form II, relaxed circular DNA. (C) DNA bending by AaHMGB1 and its truncated forms. A 32 P-labeled 123-bp DNA fragment (∼1 nM) was pre-incubated with recombinant proteins (25–50 nM) followed by ligation with T4 DNA ligase. Exonuclease III was used to verify the identity of DNA circles. The deproteinized DNA ligation products were subjected to electrophoresis on 6% non-denaturing polyacrylamide gels and visualized by autoradiography. Lm: linear multimers. Exo III, exonuclease III. These experiments were repeated three to five times each.

    Journal: PLoS ONE

    Article Title: The Dengue Vector Aedes aegypti Contains a Functional High Mobility Group Box 1 (HMGB1) Protein with a Unique Regulatory C-Terminus

    doi: 10.1371/journal.pone.0040192

    Figure Lengend Snippet: DNA transactions by recombinant AaHMGB1 proteins. (A) Preferential binding of AaHMGB1 protein to supercoiled DNA. An equimolar mixture of supercoiled and linearized plasmid pTZ19R (∼10 nM) was pre-incubated with increasing amounts of AaHMGB1 (0.5–1 µM) and the DNA–protein complexes were resolved on a 1% agarose gel, followed by staining of the gel with ethidium bromide. Form I, supercoiled DNA; L, Linear DNA; Form II, relaxed circular DNA; (B) DNA supercoiling by AaHMGB1 and its truncated forms. Circular relaxed plasmid pTZ19R DNA was incubated in the presence of topoisomerase I (Topo I) and AaHMGB1 recombinant proteins (7–14 µM). Deproteinized DNA topoisomers were resolved on 1% agarose gels, followed by staining of the gel with ethidium bromide. Form I, supercoiled DNA; Form II, relaxed circular DNA. (C) DNA bending by AaHMGB1 and its truncated forms. A 32 P-labeled 123-bp DNA fragment (∼1 nM) was pre-incubated with recombinant proteins (25–50 nM) followed by ligation with T4 DNA ligase. Exonuclease III was used to verify the identity of DNA circles. The deproteinized DNA ligation products were subjected to electrophoresis on 6% non-denaturing polyacrylamide gels and visualized by autoradiography. Lm: linear multimers. Exo III, exonuclease III. These experiments were repeated three to five times each.

    Article Snippet: Briefly, a 32 P-labeled 123-bp DNA fragment (∼1 nM) with cohesive BamHI ends were pre-incubated on ice for 20 min with appropriate amounts of recombinant proteins (25–50 nM) or total protein extracts from adult mosquitos (4 µg) in 1× T4 DNA ligase buffer (30 mM Tris–HCl, pH 7.8, 10 mM MgCl2 , 10 mM dithiothreitol, and 0.5 mM ATP; Promega) in a final volume of 20 µL.

    Techniques: Recombinant, Binding Assay, Plasmid Preparation, Incubation, Agarose Gel Electrophoresis, Staining, Labeling, Ligation, DNA Ligation, Electrophoresis, Autoradiography

    DNA bending assays by posphorylated AaHMGB1. A 32 P-labelled 123-bp DNA fragment (∼1 nM) was pre-incubated with 50 ng of AaHMGB1 that were phosphorylated by PKA (panels A and B, lanes 5 and 2, respectively) or not (panels A and B, lanes 4 and 3, respectively), or by PKC (panels C and D, lanes 5 and 2, respectively) or not (panels C and D, lanes 4 and 3, respectively), followed by ligation with T4 DNA ligase. Exonuclease III was used to verify the identity of DNA circles. The deproteinized DNA ligation products were subjected to electrophoresis on 6% non-denaturing polyacrylamide gels and visualized by autoradiography. Lm: linear multimers. These experiments were repeated five times.

    Journal: PLoS ONE

    Article Title: The Dengue Vector Aedes aegypti Contains a Functional High Mobility Group Box 1 (HMGB1) Protein with a Unique Regulatory C-Terminus

    doi: 10.1371/journal.pone.0040192

    Figure Lengend Snippet: DNA bending assays by posphorylated AaHMGB1. A 32 P-labelled 123-bp DNA fragment (∼1 nM) was pre-incubated with 50 ng of AaHMGB1 that were phosphorylated by PKA (panels A and B, lanes 5 and 2, respectively) or not (panels A and B, lanes 4 and 3, respectively), or by PKC (panels C and D, lanes 5 and 2, respectively) or not (panels C and D, lanes 4 and 3, respectively), followed by ligation with T4 DNA ligase. Exonuclease III was used to verify the identity of DNA circles. The deproteinized DNA ligation products were subjected to electrophoresis on 6% non-denaturing polyacrylamide gels and visualized by autoradiography. Lm: linear multimers. These experiments were repeated five times.

    Article Snippet: Briefly, a 32 P-labeled 123-bp DNA fragment (∼1 nM) with cohesive BamHI ends were pre-incubated on ice for 20 min with appropriate amounts of recombinant proteins (25–50 nM) or total protein extracts from adult mosquitos (4 µg) in 1× T4 DNA ligase buffer (30 mM Tris–HCl, pH 7.8, 10 mM MgCl2 , 10 mM dithiothreitol, and 0.5 mM ATP; Promega) in a final volume of 20 µL.

    Techniques: Incubation, Ligation, DNA Ligation, Electrophoresis, Autoradiography