t7 exonuclease digestion  (New England Biolabs)


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    Name:
    T7 Exonuclease
    Description:
    T7 Exonuclease 5 000 units
    Catalog Number:
    M0263L
    Price:
    261
    Category:
    Exonucleases
    Size:
    5 000 units
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    Structured Review

    New England Biolabs t7 exonuclease digestion
    T7 Exonuclease
    T7 Exonuclease 5 000 units
    https://www.bioz.com/result/t7 exonuclease digestion/product/New England Biolabs
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    t7 exonuclease digestion - by Bioz Stars, 2021-06
    95/100 stars

    Images

    1) Product Images from "Anti-apoptotic Protein BCL2 Down-regulates DNA End Joining in Cancer Cells *"

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

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M110.140350

    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.
    Figure Legend 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.

    Techniques Used: Incubation, Polyacrylamide Gel Electrophoresis

    2) Product Images from "Anti-apoptotic Protein BCL2 Down-regulates DNA End Joining in Cancer Cells *"

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

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M110.140350

    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.
    Figure Legend 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.

    Techniques Used: Incubation, Polyacrylamide Gel Electrophoresis

    Related Articles

    Avidin-Biotin Assay:

    Article Title: The structure of ends determines the pathway choice and Mre11 nuclease dependency of DNA double-strand break repair
    Article Snippet: Analysis of resection intermediates DNA intermediates were isolated from extracts by first incubating with 3 volumes of ELB buffer supplemented with 25 mM EDTA and 1/2 volume of proteinase K (10 mg/ml in H2 O) at 22°C for 2 h and then purified with the PCR purification columns following the manufacturer's protocol (Qiagen, CA, USA). .. To detect the presence of 3′ biotin on 3′ ss-overhangs or resection intermediates, the DNA was pre-incubated with ELB buffer or avidin on ice for 5 min, and then treated with Escherichia coli ExoI (NEB, MA) at 22ºC for 60 min. To analyze the intermediates of the 5′ biotin-avidin DNA, DNA was treated with E. coli ExoI (0.2 u/μl, NEB, MA) or RecJ (0.3 u/μl; NEB, MA) at 22°C for 60 min. To detect the presence of 5′ biotin, DNA was pre-incubated with ELB buffer or avidin on ice for 5 min, and then treated with T7 Exo (0.6 unit/μl; NEB, MA) at 22°C for 60 min. .. Reactions were analyzed by 1% TAE-agarose gel electrophoresis and the gels were first stained with SYBR Gold (Invitrogen, CA, USA) and then dried for exposure to film.

    Article Title: DNA double-strand breaks with 5′ adducts are efficiently channeled to the DNA2-mediated resection pathway
    Article Snippet: However, in the presence of avidin, the DNA became resistant to T7 Exo. .. Notably, the linear pBS DNA with no biotin at the end in the same reactions was equally sensitive to T7 Exo with or without avidin, indicating that it's the avidin at the 5′ end that is blocking T7 Exo. .. The DNA re-isolated after 30 min of incubation in the mock- or DNA2-depleted extracts were sensitive to T7 Exo (Figure ).

    Article Title: DNA double-strand breaks with 5′ adducts are efficiently channeled to the DNA2-mediated resection pathway
    Article Snippet: The DNA re-isolated after 30 min of incubation in the mock- or DNA2-depleted extracts were sensitive to T7 Exo (Figure ). .. Pre-incubation with avidin did not protect them from T7 Exo, suggesting that they had lost the 5′ biotin and thus avidin. ..

    other:

    Article Title: Impact of DNA ligase IV on the fidelity of end joining in human cells
    Article Snippet: We also show that DNA ligase IV–XRCC4 can contribute to the protection of ends from degradation by T7 exonuclease.

    Article Title: Anti-apoptotic Protein BCL2 Down-regulates DNA End Joining in Cancer Cells *
    Article Snippet: Interestingly, one of the circular bands (Form I) was sensitive neither to XhoI nor to T7 exonuclease, possibly because of inherent distortion of each nucleotide due to circularization of apparently a 75-mer ( B , lanes 4 and 5 ).

    Blocking Assay:

    Article Title: DNA double-strand breaks with 5′ adducts are efficiently channeled to the DNA2-mediated resection pathway
    Article Snippet: However, in the presence of avidin, the DNA became resistant to T7 Exo. .. Notably, the linear pBS DNA with no biotin at the end in the same reactions was equally sensitive to T7 Exo with or without avidin, indicating that it's the avidin at the 5′ end that is blocking T7 Exo. .. The DNA re-isolated after 30 min of incubation in the mock- or DNA2-depleted extracts were sensitive to T7 Exo (Figure ).

    Activity Assay:

    Article Title: DNA double-strand breaks with 5′ adducts are efficiently channeled to the DNA2-mediated resection pathway
    Article Snippet: To address this question, the DNA with 5′ avidin was re-isolated after incubation in the extracts and analyzed for the existence of the 5′ biotin using the bacterial T7 Exo nuclease, which degrades ds-DNA in the 5′- > 3′ direction. .. 5′ biotin per se did not inhibit the activity of T7 Exo (Figure ). .. However, in the presence of avidin, the DNA became resistant to T7 Exo.

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    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: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/t7 exonuclease digestion/product/New England Biolabs
    Average 95 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    t7 exonuclease digestion - by Bioz Stars, 2021-06
    95/100 stars
      Buy from Supplier

    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