t7 exonuclease digestion Search Results


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  • 97
    New England Biolabs t7 exonuclease digestion
    <t>T7</t> exonuclease digestion to determine the topology of end-joined products. A , diagrammatic representation of plausible end joined products by cell-free extracts. B , T7 exonuclease and XhoI digestion followed by T7 exonuclease digestion pattern of end-joined products of K562 cell-free extracts. Multiple end-joining reactions were carried out using 5′ compatible substrates, and products were incubated with either T7 exonuclease (5 units/10-μl reaction for 2 h), XhoI, or both. The products were resolved on an 8% denaturing PAGE. M , 50-nt ladder.
    T7 Exonuclease Digestion, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 97/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/t7 exonuclease digestion/product/New England Biolabs
    Average 97 stars, based on 2 article reviews
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    t7 exonuclease digestion - by Bioz Stars, 2020-08
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    92
    Thermo Fisher t7 exonuclease
    <t>T7</t> exonuclease digestion to determine the topology of end-joined products. A , diagrammatic representation of plausible end joined products by cell-free extracts. B , T7 exonuclease and XhoI digestion followed by T7 exonuclease digestion pattern of end-joined products of K562 cell-free extracts. Multiple end-joining reactions were carried out using 5′ compatible substrates, and products were incubated with either T7 exonuclease (5 units/10-μl reaction for 2 h), XhoI, or both. The products were resolved on an 8% denaturing PAGE. M , 50-nt ladder.
    T7 Exonuclease, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 38 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/t7 exonuclease/product/Thermo Fisher
    Average 92 stars, based on 38 article reviews
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    85
    GE Healthcare t7 gene 6 exonuclease digestion
    Different RecE/RecT and Redα/Redβ recombination mechanisms revealed by different substrates. ( A ) Relative recombination efficiencies achieved using RecE/RecT (ET) or Redα/Redβ (βα) expression constructs in either JC5547 or JC5519 as indicated, as a function of homology region length, are shown. Linear molecules, which were generated to contain homology regions of variable length at their ends, were allowed to recombine with a prelinearized target generated by restriction digestion. Recombination efficiencies from one experiment are shown relative to the maximum recombination efficiency of  B , which was set to 1. Recombination efficiencies are thus presented as relative and are not directly comparable to those shown in previous figures. The experiment was repeated twice, producing similar results. (█) 5547ET; (○) 5519ET; (●) 5547βα; (□) 5519βα. ( B ) Same as  A , except that the target was left intact. The maximum absolute recombination efficiency found was arbitrarily set to 1, and all other recombination efficiencies were related to this maximum. Recombination efficiencies from one experiment are shown. The experiment was repeated five times, producing similar results. The experiments shown in  A  and  B  were done in parallel with the same batches of competent cells and PCR fragments. Colony numbers observed in these experiments were adjusted to relative recombination efficiencies by relating to the maximums obtained in  B , which were set to 1. (█) 5547ET; (○) 5519ET; (●) 5547βα; (□) 5519βα. ( C ) Recombination efficiencies in the presence of the indicated expression constructs in JC5519 using two linear molecules, both of which were preresected in vitro using T7 gene six exonuclease. Data from one representative experiment is shown, in which pools of preresected molecules (which shared 400 nucleotide homology regions and were generated using six different incubation conditions with T7 gene 6 exonuclease) were recombined. C, no exogenous protein expressed; β, expression of Redβ only; βα, coexpression of Redβ and Redα; T, expression of RecT only; ET, coexpression of RecE and RecT. Recombination efficiencies are shown relative to the maximum of  B .
    T7 Gene 6 Exonuclease Digestion, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 85/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    New England Biolabs exonuclease t7
    Dual biotin-labeled DNA fragments form a closed substrate on the surface of the BLI probe. (A) A schematic of a double digestion assay using <t>Exonuclease</t> T7 + Exonuclease VII and PCR. PCR was performed using M13F, M13R oligos, and DNA attached to the BLI surface as a template. (B) The BLI probe was severed from the plastic adaptor and immerged into a PCR master mix. (C) Dual biotin-labeled DNA fragments on the BLI surface were resistant to Exo T7 + Exo VII digestion while single biotin-labeled DNA fragments on the BLI surface were not.
    Exonuclease T7, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    New England Biolabs bacteriophage t7 gene6 exonuclease
    Dual biotin-labeled DNA fragments form a closed substrate on the surface of the BLI probe. (A) A schematic of a double digestion assay using <t>Exonuclease</t> T7 + Exonuclease VII and PCR. PCR was performed using M13F, M13R oligos, and DNA attached to the BLI surface as a template. (B) The BLI probe was severed from the plastic adaptor and immerged into a PCR master mix. (C) Dual biotin-labeled DNA fragments on the BLI surface were resistant to Exo T7 + Exo VII digestion while single biotin-labeled DNA fragments on the BLI surface were not.
    Bacteriophage T7 Gene6 Exonuclease, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 91/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/bacteriophage t7 gene6 exonuclease/product/New England Biolabs
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    Image Search Results


    T7 exonuclease digestion to determine the topology of end-joined products. A , diagrammatic representation of plausible end joined products by cell-free extracts. B , T7 exonuclease and XhoI digestion followed by T7 exonuclease digestion pattern of end-joined products of K562 cell-free extracts. Multiple end-joining reactions were carried out using 5′ compatible substrates, and products were incubated with either T7 exonuclease (5 units/10-μl reaction for 2 h), XhoI, or both. The products were resolved on an 8% denaturing PAGE. M , 50-nt ladder.

    Journal: The Journal of Biological Chemistry

    Article Title: Anti-apoptotic Protein BCL2 Down-regulates DNA End Joining in Cancer Cells *

    doi: 10.1074/jbc.M110.140350

    Figure Lengend Snippet: T7 exonuclease digestion to determine the topology of end-joined products. A , diagrammatic representation of plausible end joined products by cell-free extracts. B , T7 exonuclease and XhoI digestion followed by T7 exonuclease digestion pattern of end-joined products of K562 cell-free extracts. Multiple end-joining reactions were carried out using 5′ compatible substrates, and products were incubated with either T7 exonuclease (5 units/10-μl reaction for 2 h), XhoI, or both. The products were resolved on an 8% denaturing PAGE. M , 50-nt ladder.

    Article Snippet: T7 exonuclease digestion was performed by incubating purified EJ products with either increasing concentrations or 5 units of T7 exonuclease (New England Biolabs) at 25 °C for 2 h. In some cases, a fraction of EJ products was digested with XhoI (4 units) (37 °C for 4 h) prior to T7 exonuclease digestion.

    Techniques: Incubation, Polyacrylamide Gel Electrophoresis

    Determination of the frequency of DNA looping by enhancement of ligase-catalyzed DNA circularization. A , schematic depiction of the circularization assay showing the pUC19-based substrate that contained γ-γ or α-γ iterons cloned at the indicated locations on either side of a unique EcoR1 site. A similar plasmid having a single γ iteron on one side and the 7 γ iterons on the other was also constructed. The assay steps including linearization by EcoR1, 5′-end labeling, incubation with either π cop, or with a 1:1 mixture of wt π and π cop, ligation with T4 DNA ligase at 16 °C for various periods of time, and digestion with T7 gene 6 exonuclease are shown; 20 fmol of DNA were incubated with a total of 128 pmol of π.  B , DNA circularization kinetics showing that there was a small difference between γ-γ and γ-7 γ iteron interactions; the control experiment with a solo γ iteron is also shown.  C , DNA circularization kinetics with various combinations of DNA substrates and π proteins as indicated. The α-γ plasmid incubated with a 1:1 mixture of wt π and π cop had a small but distinct and reproducible enhancement in ligation kinetics over the same incubated with π cop only. The γ-γ plasmid incubated with π monomer-dimer mixture had a much higher rate of DNA circularization over that when only π cop was used. Data were collected from six separate sets of experiments and are presented with the S.D. as  error bars. D , ligation experiments showing that π D226A (non-DNA binding mutant dimers) did not complement π cop to loop two γ-γ iterons. Controls show that under identical conditions, wt π complemented π cop in promoting γ-γ interaction, and there was no interaction as expected when π was withheld from the reaction.

    Journal: The Journal of Biological Chemistry

    Article Title: Investigations of ? Initiator Protein-mediated Interaction between Replication Origins ? and ? of the Plasmid R6K *

    doi: 10.1074/jbc.M109.067439

    Figure Lengend Snippet: Determination of the frequency of DNA looping by enhancement of ligase-catalyzed DNA circularization. A , schematic depiction of the circularization assay showing the pUC19-based substrate that contained γ-γ or α-γ iterons cloned at the indicated locations on either side of a unique EcoR1 site. A similar plasmid having a single γ iteron on one side and the 7 γ iterons on the other was also constructed. The assay steps including linearization by EcoR1, 5′-end labeling, incubation with either π cop, or with a 1:1 mixture of wt π and π cop, ligation with T4 DNA ligase at 16 °C for various periods of time, and digestion with T7 gene 6 exonuclease are shown; 20 fmol of DNA were incubated with a total of 128 pmol of π. B , DNA circularization kinetics showing that there was a small difference between γ-γ and γ-7 γ iteron interactions; the control experiment with a solo γ iteron is also shown. C , DNA circularization kinetics with various combinations of DNA substrates and π proteins as indicated. The α-γ plasmid incubated with a 1:1 mixture of wt π and π cop had a small but distinct and reproducible enhancement in ligation kinetics over the same incubated with π cop only. The γ-γ plasmid incubated with π monomer-dimer mixture had a much higher rate of DNA circularization over that when only π cop was used. Data were collected from six separate sets of experiments and are presented with the S.D. as error bars. D , ligation experiments showing that π D226A (non-DNA binding mutant dimers) did not complement π cop to loop two γ-γ iterons. Controls show that under identical conditions, wt π complemented π cop in promoting γ-γ interaction, and there was no interaction as expected when π was withheld from the reaction.

    Article Snippet: The reaction products were digested with T7 exonuclease, and measurements of π-mediated enhancement of circle formation were carried out by counting the exonuclease-resistant label.

    Techniques: Clone Assay, Plasmid Preparation, Construct, End Labeling, Incubation, Ligation, Binding Assay, Mutagenesis

    Different RecE/RecT and Redα/Redβ recombination mechanisms revealed by different substrates. ( A ) Relative recombination efficiencies achieved using RecE/RecT (ET) or Redα/Redβ (βα) expression constructs in either JC5547 or JC5519 as indicated, as a function of homology region length, are shown. Linear molecules, which were generated to contain homology regions of variable length at their ends, were allowed to recombine with a prelinearized target generated by restriction digestion. Recombination efficiencies from one experiment are shown relative to the maximum recombination efficiency of  B , which was set to 1. Recombination efficiencies are thus presented as relative and are not directly comparable to those shown in previous figures. The experiment was repeated twice, producing similar results. (█) 5547ET; (○) 5519ET; (●) 5547βα; (□) 5519βα. ( B ) Same as  A , except that the target was left intact. The maximum absolute recombination efficiency found was arbitrarily set to 1, and all other recombination efficiencies were related to this maximum. Recombination efficiencies from one experiment are shown. The experiment was repeated five times, producing similar results. The experiments shown in  A  and  B  were done in parallel with the same batches of competent cells and PCR fragments. Colony numbers observed in these experiments were adjusted to relative recombination efficiencies by relating to the maximums obtained in  B , which were set to 1. (█) 5547ET; (○) 5519ET; (●) 5547βα; (□) 5519βα. ( C ) Recombination efficiencies in the presence of the indicated expression constructs in JC5519 using two linear molecules, both of which were preresected in vitro using T7 gene six exonuclease. Data from one representative experiment is shown, in which pools of preresected molecules (which shared 400 nucleotide homology regions and were generated using six different incubation conditions with T7 gene 6 exonuclease) were recombined. C, no exogenous protein expressed; β, expression of Redβ only; βα, coexpression of Redβ and Redα; T, expression of RecT only; ET, coexpression of RecE and RecT. Recombination efficiencies are shown relative to the maximum of  B .

    Journal: Genes & Development

    Article Title: RecE/RecT and Red?/Red? initiate double-stranded break repair by specifically interacting with their respective partners

    doi:

    Figure Lengend Snippet: Different RecE/RecT and Redα/Redβ recombination mechanisms revealed by different substrates. ( A ) Relative recombination efficiencies achieved using RecE/RecT (ET) or Redα/Redβ (βα) expression constructs in either JC5547 or JC5519 as indicated, as a function of homology region length, are shown. Linear molecules, which were generated to contain homology regions of variable length at their ends, were allowed to recombine with a prelinearized target generated by restriction digestion. Recombination efficiencies from one experiment are shown relative to the maximum recombination efficiency of B , which was set to 1. Recombination efficiencies are thus presented as relative and are not directly comparable to those shown in previous figures. The experiment was repeated twice, producing similar results. (█) 5547ET; (○) 5519ET; (●) 5547βα; (□) 5519βα. ( B ) Same as A , except that the target was left intact. The maximum absolute recombination efficiency found was arbitrarily set to 1, and all other recombination efficiencies were related to this maximum. Recombination efficiencies from one experiment are shown. The experiment was repeated five times, producing similar results. The experiments shown in A and B were done in parallel with the same batches of competent cells and PCR fragments. Colony numbers observed in these experiments were adjusted to relative recombination efficiencies by relating to the maximums obtained in B , which were set to 1. (█) 5547ET; (○) 5519ET; (●) 5547βα; (□) 5519βα. ( C ) Recombination efficiencies in the presence of the indicated expression constructs in JC5519 using two linear molecules, both of which were preresected in vitro using T7 gene six exonuclease. Data from one representative experiment is shown, in which pools of preresected molecules (which shared 400 nucleotide homology regions and were generated using six different incubation conditions with T7 gene 6 exonuclease) were recombined. C, no exogenous protein expressed; β, expression of Redβ only; βα, coexpression of Redβ and Redα; T, expression of RecT only; ET, coexpression of RecE and RecT. Recombination efficiencies are shown relative to the maximum of B .

    Article Snippet: T7 gene 6 exonuclease digestion was done in the buffer provided by the supplier (Amersham, Inc.), using ∼1.5 μg linear substrate per reaction.

    Techniques: Expressing, Construct, Generated, Polymerase Chain Reaction, In Vitro, Incubation

    Dual biotin-labeled DNA fragments form a closed substrate on the surface of the BLI probe. (A) A schematic of a double digestion assay using Exonuclease T7 + Exonuclease VII and PCR. PCR was performed using M13F, M13R oligos, and DNA attached to the BLI surface as a template. (B) The BLI probe was severed from the plastic adaptor and immerged into a PCR master mix. (C) Dual biotin-labeled DNA fragments on the BLI surface were resistant to Exo T7 + Exo VII digestion while single biotin-labeled DNA fragments on the BLI surface were not.

    Journal: bioRxiv

    Article Title: ParB spreading on DNA requires cytidine triphosphate in vitro

    doi: 10.1101/2019.12.11.865972

    Figure Lengend Snippet: Dual biotin-labeled DNA fragments form a closed substrate on the surface of the BLI probe. (A) A schematic of a double digestion assay using Exonuclease T7 + Exonuclease VII and PCR. PCR was performed using M13F, M13R oligos, and DNA attached to the BLI surface as a template. (B) The BLI probe was severed from the plastic adaptor and immerged into a PCR master mix. (C) Dual biotin-labeled DNA fragments on the BLI surface were resistant to Exo T7 + Exo VII digestion while single biotin-labeled DNA fragments on the BLI surface were not.

    Article Snippet: To verify that dual biotin-labeled DNA fragments formed a closed substrate on the surface of the BLI probe, we performed a double digestion with Exonuclease T7 and Exonuclease VII (NEB) ( ).

    Techniques: Labeling, Polymerase Chain Reaction