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  • 94
    New England Biolabs i rfi circular dsdna
    Substrate specificity of Endo IV. The activity of Endo IV (1, 0.5, 0.2, 0.1 or 0.05 μg/ml) was assayed with phiX174 <t>RFI</t> circular <t>dsDNA</t> ( A ), phiX174 RFI linear dsDNA ( B ), phiX174 virion circular ssDNA ( C ), linear ssRNA used for the in vitro synthesis of the GST-Endo IV fusion protein ( D ), heat-denatured T4 genomic dsDNA containing gluc-dHMC ( E ), or heat-denatured dC-substituted T4 (T4dC) genomic dsDNA ( F ) as the substrate (5 μg/ml). The reaction products were separated by electrophoresis on a 1.0% agarose gel and stained with SYBR Gold. Lanes M and (–) contain a 1 kb DNA ladder and a reaction mixture incubated in the absence of the enzyme, respectively.
    I Rfi Circular Dsdna, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 94/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/i rfi circular dsdna/product/New England Biolabs
    Average 94 stars, based on 7 article reviews
    Price from $9.99 to $1999.99
    i rfi circular dsdna - by Bioz Stars, 2020-08
    94/100 stars
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    85
    Thermo Fisher i rfi dna
    MoMLV PIC is compacted by BAF. (A) Sedimentation properties of initial (panel 1), salt-stripped (panel 2), BAF-reconstituted (panel 3), HMGA1-reconstituted (panels 4 and 5), and HMGB1-reconstituted (panels 6 and 7) PICs. The MoMLV PIC fraction was salt stripped with 750 mM KCl and then incubated with BAF, HMGA1, or HMGB1 protein in the presence of 400 mM KCl (panels 1, 2, 3, 4, and 6) or 150 mM KCl (panels 5 and 7). After gel filtration, each reconstituted PIC fraction was layered on a 15 to 30% sucrose gradient and centrifuged at 30,000 rpm for 1 h with a Beckman rotor. The gradient was fractionated from the top to bottom into 22 fractions, and viral <t>DNA</t> in each fraction was detected by Southern blotting. (B) Intermolecular integration activities of initial, salt-stripped, and reconstituted PICs. Salt-stripped PICs were incubated with BAF (lane 3), HMGA1 (lanes 4 and 6), or HMGB1 (lanes 5 and 7) in the presence of 400 mM KCl (lanes 1 to 5) or 150 mM KCl (lanes 6 and 7) and then added to the integration reaction mixture containing φX174 <t>RFI</t> DNA as the target DNA to check integration activities. After incubation, DNA products from the reaction were digested with Bam HI and detected by Southern blotting with a 32 ).
    I Rfi Dna, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/i rfi dna/product/Thermo Fisher
    Average 85 stars, based on 7 article reviews
    Price from $9.99 to $1999.99
    i rfi dna - by Bioz Stars, 2020-08
    85/100 stars
      Buy from Supplier

    Image Search Results


    Substrate specificity of Endo IV. The activity of Endo IV (1, 0.5, 0.2, 0.1 or 0.05 μg/ml) was assayed with phiX174 RFI circular dsDNA ( A ), phiX174 RFI linear dsDNA ( B ), phiX174 virion circular ssDNA ( C ), linear ssRNA used for the in vitro synthesis of the GST-Endo IV fusion protein ( D ), heat-denatured T4 genomic dsDNA containing gluc-dHMC ( E ), or heat-denatured dC-substituted T4 (T4dC) genomic dsDNA ( F ) as the substrate (5 μg/ml). The reaction products were separated by electrophoresis on a 1.0% agarose gel and stained with SYBR Gold. Lanes M and (–) contain a 1 kb DNA ladder and a reaction mixture incubated in the absence of the enzyme, respectively.

    Journal: Nucleic Acids Research

    Article Title: Biochemical analysis of the substrate specificity and sequence preference of endonuclease IV from bacteriophage T4, a dC-specific endonuclease implicated in restriction of dC-substituted T4 DNA synthesis

    doi: 10.1093/nar/gkl553

    Figure Lengend Snippet: Substrate specificity of Endo IV. The activity of Endo IV (1, 0.5, 0.2, 0.1 or 0.05 μg/ml) was assayed with phiX174 RFI circular dsDNA ( A ), phiX174 RFI linear dsDNA ( B ), phiX174 virion circular ssDNA ( C ), linear ssRNA used for the in vitro synthesis of the GST-Endo IV fusion protein ( D ), heat-denatured T4 genomic dsDNA containing gluc-dHMC ( E ), or heat-denatured dC-substituted T4 (T4dC) genomic dsDNA ( F ) as the substrate (5 μg/ml). The reaction products were separated by electrophoresis on a 1.0% agarose gel and stained with SYBR Gold. Lanes M and (–) contain a 1 kb DNA ladder and a reaction mixture incubated in the absence of the enzyme, respectively.

    Article Snippet: Purified Endo IV was diluted in a solution containing 10 mM Tris–HCl (pH 8.0), 1 mM DTT and BSA (0.1 mg/ml) and was included in the reaction mixture in a volume of 1 μl. phiX174 replicative form I (RFI) circular dsDNA (New England BioLabs), phiX174 RFI linear dsDNA prepared by PstI digestion, phiX174 virion circular ssDNA (New England BioLabs), linear ssRNA used for synthesis of the GST-Endo IV fusion protein, and heat-denatured T4 or T4dC genomic dsDNA were used as substrates at a final concentration of 5 μg/ml.

    Techniques: Activity Assay, In Vitro, Electrophoresis, Agarose Gel Electrophoresis, Staining, Incubation

    MoMLV PIC is compacted by BAF. (A) Sedimentation properties of initial (panel 1), salt-stripped (panel 2), BAF-reconstituted (panel 3), HMGA1-reconstituted (panels 4 and 5), and HMGB1-reconstituted (panels 6 and 7) PICs. The MoMLV PIC fraction was salt stripped with 750 mM KCl and then incubated with BAF, HMGA1, or HMGB1 protein in the presence of 400 mM KCl (panels 1, 2, 3, 4, and 6) or 150 mM KCl (panels 5 and 7). After gel filtration, each reconstituted PIC fraction was layered on a 15 to 30% sucrose gradient and centrifuged at 30,000 rpm for 1 h with a Beckman rotor. The gradient was fractionated from the top to bottom into 22 fractions, and viral DNA in each fraction was detected by Southern blotting. (B) Intermolecular integration activities of initial, salt-stripped, and reconstituted PICs. Salt-stripped PICs were incubated with BAF (lane 3), HMGA1 (lanes 4 and 6), or HMGB1 (lanes 5 and 7) in the presence of 400 mM KCl (lanes 1 to 5) or 150 mM KCl (lanes 6 and 7) and then added to the integration reaction mixture containing φX174 RFI DNA as the target DNA to check integration activities. After incubation, DNA products from the reaction were digested with Bam HI and detected by Southern blotting with a 32 ).

    Journal: Journal of Virology

    Article Title: Regulatory Mechanisms by Which Barrier-to-Autointegration Factor Blocks Autointegration and Stimulates Intermolecular Integration of Moloney Murine Leukemia Virus Preintegration Complexes

    doi: 10.1128/JVI.76.23.12376-12380.2002

    Figure Lengend Snippet: MoMLV PIC is compacted by BAF. (A) Sedimentation properties of initial (panel 1), salt-stripped (panel 2), BAF-reconstituted (panel 3), HMGA1-reconstituted (panels 4 and 5), and HMGB1-reconstituted (panels 6 and 7) PICs. The MoMLV PIC fraction was salt stripped with 750 mM KCl and then incubated with BAF, HMGA1, or HMGB1 protein in the presence of 400 mM KCl (panels 1, 2, 3, 4, and 6) or 150 mM KCl (panels 5 and 7). After gel filtration, each reconstituted PIC fraction was layered on a 15 to 30% sucrose gradient and centrifuged at 30,000 rpm for 1 h with a Beckman rotor. The gradient was fractionated from the top to bottom into 22 fractions, and viral DNA in each fraction was detected by Southern blotting. (B) Intermolecular integration activities of initial, salt-stripped, and reconstituted PICs. Salt-stripped PICs were incubated with BAF (lane 3), HMGA1 (lanes 4 and 6), or HMGB1 (lanes 5 and 7) in the presence of 400 mM KCl (lanes 1 to 5) or 150 mM KCl (lanes 6 and 7) and then added to the integration reaction mixture containing φX174 RFI DNA as the target DNA to check integration activities. After incubation, DNA products from the reaction were digested with Bam HI and detected by Southern blotting with a 32 ).

    Article Snippet: Integration activities of reconstituted PICs were evaluated by the previously described integration activity assay with φX174 replicative form I (RFI) DNA (Gibco BRL) as the target DNA ( ).

    Techniques: Sedimentation, Incubation, Filtration, Southern Blot