exonuclease vii  (Thermo Fisher)


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    Name:
    Exonuclease VII
    Description:
    Exonuclease VII is a strict single-strand directed enzyme with 5'→3' and 3'→5' exonuclease activities, making it the only bi-directional exonuclease with single-stranded specificity. Exonuclease VII has no apparent requirement for divalent cations, as it is fully active in the presence of EDTA. Initial reaction products are acid insoluble oligonucleotides which are further hydrolyzed into acid soluble form. The products of limited digestions are small oligomers (dimers to dodecamers). Properties: Molecular Weight: xseA subunit = 51.8 kDa; xseB subunit = 8.8 kDa Optimum Temperature: 37 °C Optimum pH: 8.0 Inactivation: 95 °C for 10 min. Purity: Greater than 95% pure as determined by SDS-PAGE. Tested for contaminating endonucleases, double-stranded exonucleases, and ribonucleases. Storage Buffer: 50 mM Tris-HCl (pH 8.0), 200 mM NaCl, 0.5 mM EDTA, 10 mM DTT, 50% glycerol. Assay Conditions: The reaction (50 µL) contains 50 mM Tris-HCl (pH 7.9), 50 mM potassium phosphate (pH 7.6), 8.3 mM EDTA, 10 mM 2-mercaptoethanol, denatured DNA, and enzyme. Unit Definition: One unit is the amount of enzyme required to catalyze the conversion of 1 nmol of nucleotide to acid-soluble nucleotide in 30 min at 37 °C under standard assay conditions. Concentration: 10 units/µL Source: E. coli strain containing overproducing clones encoding both the large (xseA) and small (xseB) subunits of Exonuclease VII. Protocol: Typical Reaction Conditions (50 µL) 70 mM Tris-HCI, pH 8.0 8 mM EDTA 10 mM 2-mercaptoethanol 1 µg DNA 50 µg/mL 0.2 units Exonuclease VII Incubate at 37 °C for 30 min. Inactivate the enzyme by heating to 95 °C for 10 min. Note that Exonuclease VII is not inhibited by EDTA. Note: A typical dilution buffer for use with Exonuclease VII is 50 mM Tris, pH 7.9, 1 mM DTT and 0.5 mg/mL acetylated BSA.Applications:   1. Mapping positions of introns in genomic DNA.   2. Removal of vector DNA from inserts with poly (dA-T) tails.   3. Removal of excess PCR primers.   4. Removal of single-stranded over-hangs produced by restriction endonucleases.
    Catalog Number:
    70082Z1000UN
    Price:
    None
    Applications:
    PCR|PCR & Real-Time PCR|Sequencing
    Size:
    1000 units
    Category:
    Proteins, Enzymes, & Peptides, PCR & Cloning Enzymes, DNA⁄RNA Modifying Enzymes
    Score:
    85
    Buy from Supplier


    Structured Review

    Thermo Fisher exonuclease vii
    Exonuclease <t>VII</t> protection of product DNA resolves roles of the TERT ring RNP and TEN domain in DNA handling
    Exonuclease VII is a strict single-strand directed enzyme with 5'→3' and 3'→5' exonuclease activities, making it the only bi-directional exonuclease with single-stranded specificity. Exonuclease VII has no apparent requirement for divalent cations, as it is fully active in the presence of EDTA. Initial reaction products are acid insoluble oligonucleotides which are further hydrolyzed into acid soluble form. The products of limited digestions are small oligomers (dimers to dodecamers). Properties: Molecular Weight: xseA subunit = 51.8 kDa; xseB subunit = 8.8 kDa Optimum Temperature: 37 °C Optimum pH: 8.0 Inactivation: 95 °C for 10 min. Purity: Greater than 95% pure as determined by SDS-PAGE. Tested for contaminating endonucleases, double-stranded exonucleases, and ribonucleases. Storage Buffer: 50 mM Tris-HCl (pH 8.0), 200 mM NaCl, 0.5 mM EDTA, 10 mM DTT, 50% glycerol. Assay Conditions: The reaction (50 µL) contains 50 mM Tris-HCl (pH 7.9), 50 mM potassium phosphate (pH 7.6), 8.3 mM EDTA, 10 mM 2-mercaptoethanol, denatured DNA, and enzyme. Unit Definition: One unit is the amount of enzyme required to catalyze the conversion of 1 nmol of nucleotide to acid-soluble nucleotide in 30 min at 37 °C under standard assay conditions. Concentration: 10 units/µL Source: E. coli strain containing overproducing clones encoding both the large (xseA) and small (xseB) subunits of Exonuclease VII. Protocol: Typical Reaction Conditions (50 µL) 70 mM Tris-HCI, pH 8.0 8 mM EDTA 10 mM 2-mercaptoethanol 1 µg DNA 50 µg/mL 0.2 units Exonuclease VII Incubate at 37 °C for 30 min. Inactivate the enzyme by heating to 95 °C for 10 min. Note that Exonuclease VII is not inhibited by EDTA. Note: A typical dilution buffer for use with Exonuclease VII is 50 mM Tris, pH 7.9, 1 mM DTT and 0.5 mg/mL acetylated BSA.Applications:   1. Mapping positions of introns in genomic DNA.   2. Removal of vector DNA from inserts with poly (dA-T) tails.   3. Removal of excess PCR primers.   4. Removal of single-stranded over-hangs produced by restriction endonucleases.
    https://www.bioz.com/result/exonuclease vii/product/Thermo Fisher
    Average 89 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    exonuclease vii - by Bioz Stars, 2019-10
    89/100 stars

    Images

    1) Product Images from "Human telomerase specialization for repeat synthesis by unique handling of primer-template duplex"

    Article Title: Human telomerase specialization for repeat synthesis by unique handling of primer-template duplex

    Journal:

    doi: 10.1002/embj.201387205

    Exonuclease VII protection of product DNA resolves roles of the TERT ring RNP and TEN domain in DNA handling
    Figure Legend Snippet: Exonuclease VII protection of product DNA resolves roles of the TERT ring RNP and TEN domain in DNA handling

    Techniques Used:

    Related Articles

    Clone Assay:

    Article Title: Functional Analysis of Three Arabidopsis ARGONAUTES Using Slicer-Defective Mutants
    Article Snippet: Ligation reactions were incubated overnight at 16°C. .. Digestion of unligated Cloning Linker 1 was done by first adding 20 units of deadenylase (NEB) to the overnight ligation and incubating at 30°C for 15 min, then adding 10 units of Exonuclease VII (Affymetrix) and incubating at 37°C for 15 min. Next, 15 pmol of RNA 5′ adapter (see online) was denatured at 70°C for 2 min followed by transfer to ice for at least 2 min. Denatured RNA 5′ adapter was added to the ligation mixture with 0.48 mM of ATP and five units of T4 RNA Ligase 1 (Ambion), and the resulting mixture was incubated at 28°C for 1 h, then placed on ice. .. For cDNA synthesis, 5 µM of RT primer (see online) was used with the SuperScript III first-strand synthesis system (Invitrogen).

    Incubation:

    Article Title: Human telomerase specialization for repeat synthesis by unique handling of primer-template duplex
    Article Snippet: The primer extension reaction was initiated by the addition, to final concentration, of 50 mM Tris-acetate at pH 8, 3 mM MgCl2 , 1 mM EGTA, 1 mM spermidine, 5 mM DTT, 5% glycerol, 0.1 μM α-32 P dGTP and, if included, 250 μM of other dNTPs and/or 500 μM of ddNTPs in a final volume of 20 μl. .. The reaction mixtures were incubated at room temperature for 1 min, followed by the addition of 5 units of exonuclease VII (ExoVII, Affymetrix). .. The reaction mixtures were incubated at room temperature for 5 min, then stopped by the addition of 80 μl stop buffer (50 mM Tris–HCl at pH 7.9, 20 mM EDTA, and 0.2% SDS).

    Article Title: DNA‐binding determinants and cellular thresholds for human telomerase repeat addition processivity
    Article Snippet: Telomerase immobilized on FLAG‐antibody resin was allowed to bind 5′‐T21 GT2 AG2 ‐3′ primer supplied at 1 μM and incubated at room temperature for 30 min. Primer extension was initiated by addition of reaction buffer and a final concentration of 0.1 μM α‐32 P dGTP and, if included, 250 μM dATP, 250 μM dTTP, and/or 500 μM of ddNTP. .. Reactions were incubated at room temperature for 1 min, followed by addition of five units of ExoVII (Affymetrix). .. 32 P end‐labeled oligonucleotides were loaded as size markers.

    Article Title: Alternative splicing enriched cDNA libraries identify breast cancer-associated transcripts
    Article Snippet: Double-stranded cDNA molecules were heated at 96°C for 20 min and incubated at 42°C for 24 hours in a mixture of 0.2% SDS, 0.5 M NaCl, 0.05 M Tris-HCl pH 7.5 and 30% formamide. .. Exonuclease VII (USB Corporation) cleavage was performed in 70 mM Tris-HCI, pH 8.0; 8 mM EDTA, pH 8.0; 10 mM 2-mercaptoethanol; 50 µg/ml BSA and 0.2 U of the enzyme and incubated at 37°C for 30 min. .. Fifteen units of the restriction enzyme II (New England Biolabs) was used for each 500 ng of cDNA in 1X buffer.

    Article Title: Functional Analysis of Three Arabidopsis ARGONAUTES Using Slicer-Defective Mutants
    Article Snippet: Ligation reactions were incubated overnight at 16°C. .. Digestion of unligated Cloning Linker 1 was done by first adding 20 units of deadenylase (NEB) to the overnight ligation and incubating at 30°C for 15 min, then adding 10 units of Exonuclease VII (Affymetrix) and incubating at 37°C for 15 min. Next, 15 pmol of RNA 5′ adapter (see online) was denatured at 70°C for 2 min followed by transfer to ice for at least 2 min. Denatured RNA 5′ adapter was added to the ligation mixture with 0.48 mM of ATP and five units of T4 RNA Ligase 1 (Ambion), and the resulting mixture was incubated at 28°C for 1 h, then placed on ice. .. For cDNA synthesis, 5 µM of RT primer (see online) was used with the SuperScript III first-strand synthesis system (Invitrogen).

    Article Title: Human Immunodeficiency Virus Type 1 Nucleocapsid Protein Can Prevent Self-Priming of Minus-Strand Strong Stop DNA by Promoting the Annealing of Short Oligonucleotides to Hairpin Sequences
    Article Snippet: After 20 min at 37°C, the NC-oligomer reactions were mixed and incubated at 37°C for 8 min. T4 DNA ligase (New England Biolabs) was added in 50-fold molar excess, and the reactions were incubated at 22°C for 30 min. Ligase was inactivated by heating to 65°C, and the reactions were placed on ice. .. Then 0.3 U of exonuclease VII (Exo VII; Gibco/BRL, Rockville, Md.) was added, and the mixture was incubated for 2 h at 37°C. .. Reactions were stopped by the addition of an equal volume of formamide loading dye containing 0.1% SDS and fractionated by electrophoresis on a 12% denaturing acrylamide gel.

    Expressing:

    Article Title: A random six-phase switch regulates pneumococcal virulence via global epigenetic changes
    Article Snippet: Incompletely formed SMRTbell templates were digested using Exonuclease III (New England Biolabs) and Exonuclease VII (Affymetrix, OH, USA). .. Methylation sites of variants SpnIIIA-C were experimentally confirmed by protection of pDP28 DNA from digestion by methylation sensitive enzymes with overlapping target specificity.

    Genome Wide:

    Article Title: Identification of a Pseudomonas aeruginosa PAO1 DNA Methyltransferase, Its Targets, and Physiological Roles
    Article Snippet: A mixture of exonuclease III and exonuclease VII (Affymetrix, High Wycombe, United Kingdom) was utilized. .. A mixture of exonuclease III and exonuclease VII (Affymetrix, High Wycombe, United Kingdom) was utilized.

    Recombinase Polymerase Amplification:

    Article Title: The RecA/RAD51 protein drives migration of Holliday junctions via polymerization on DNA
    Article Snippet: Human RAD51, RAD51K133R , RAD51K133A , and RPA were purified as described ( ). .. SSB, Shrimp alkaline phosphatase, and Exonuclease VII were from USB Corp., and Terminal deoxynucleotidyl transferase, restriction endonucleases, Exonuclease I, and T4 polynucleotide kinase were from New England Biolabs.

    Ligation:

    Article Title: Functional Analysis of Three Arabidopsis ARGONAUTES Using Slicer-Defective Mutants
    Article Snippet: Ligation reactions were incubated overnight at 16°C. .. Digestion of unligated Cloning Linker 1 was done by first adding 20 units of deadenylase (NEB) to the overnight ligation and incubating at 30°C for 15 min, then adding 10 units of Exonuclease VII (Affymetrix) and incubating at 37°C for 15 min. Next, 15 pmol of RNA 5′ adapter (see online) was denatured at 70°C for 2 min followed by transfer to ice for at least 2 min. Denatured RNA 5′ adapter was added to the ligation mixture with 0.48 mM of ATP and five units of T4 RNA Ligase 1 (Ambion), and the resulting mixture was incubated at 28°C for 1 h, then placed on ice. .. For cDNA synthesis, 5 µM of RT primer (see online) was used with the SuperScript III first-strand synthesis system (Invitrogen).

    Footprinting:

    Article Title: Human telomerase specialization for repeat synthesis by unique handling of primer-template duplex
    Article Snippet: The reaction mixtures were incubated at room temperature for 1 min, followed by the addition of 5 units of exonuclease VII (ExoVII, Affymetrix). .. The reaction mixtures were incubated at room temperature for 1 min, followed by the addition of 5 units of exonuclease VII (ExoVII, Affymetrix).

    Article Title: DNA‐binding determinants and cellular thresholds for human telomerase repeat addition processivity
    Article Snippet: Reactions were incubated at room temperature for 1 min, followed by addition of five units of ExoVII (Affymetrix). .. Reactions were incubated at room temperature for 1 min, followed by addition of five units of ExoVII (Affymetrix).

    DNA Sequencing:

    Article Title: First genome sequences of Achromobacter phages reveal new members of the N4 family
    Article Snippet: A mixture of Exonuclease III and Exonuclease VII (Affymetrix, High Wycombe, UK) was utilized. .. Conditions for annealing of sequencing primers and binding of polymerase to purified SMRTbell™ templates were assessed with the Calculator in RS Remote, PacificBiosciences, Menlo Park, CA, USA.

    Sequencing:

    Article Title: Identification of a Pseudomonas aeruginosa PAO1 DNA Methyltransferase, Its Targets, and Physiological Roles
    Article Snippet: Paragraph title: SMRT sequencing and motif search. ... A mixture of exonuclease III and exonuclease VII (Affymetrix, High Wycombe, United Kingdom) was utilized.

    Article Title: A role for the bacterial GATC methylome in antibiotic stress survival
    Article Snippet: Paragraph title: Genomic DNA extraction and PacBio sequencing ... Incompletely formed SMRTbell templates were digested using Exonuclease III (New England Biolabs) and Exonuclease VII (Affymetrix).

    Article Title: First genome sequences of Achromobacter phages reveal new members of the N4 family
    Article Snippet: Paragraph title: Sample preparation and sequencing ... A mixture of Exonuclease III and Exonuclease VII (Affymetrix, High Wycombe, UK) was utilized.

    Article Title: Specificity of the ModA11, ModA12 and ModD1 epigenetic regulator N6-adenine DNA methyltransferases of Neisseria meningitidis
    Article Snippet: Paragraph title: Methylome determination by SMRT sequencing ... Incompletely formed SMRTbell templates were digested using Exonuclease III (NEB) and Exonuclease VII (Affymetrix; Cleveland, OH, USA).

    Article Title: A random six-phase switch regulates pneumococcal virulence via global epigenetic changes
    Article Snippet: Incompletely formed SMRTbell templates were digested using Exonuclease III (New England Biolabs) and Exonuclease VII (Affymetrix, OH, USA). .. Methylation sites of variants SpnIIIA-C were experimentally confirmed by protection of pDP28 DNA from digestion by methylation sensitive enzymes with overlapping target specificity.

    Article Title: Methylomic and phenotypic analysis of the ModH5 phasevarion of Helicobacter pylori
    Article Snippet: Paragraph title: SMRT sequencing ... Incompletely formed SMRTbell templates were digested using Exonuclease III (NEB) and Exonuclease VII (Affymetrix; Cleveland, OH, USA).

    Article Title: Genome Annotation Provides Insight into Carbon Monoxide and Hydrogen Metabolism in Rubrivivax gelatinosus
    Article Snippet: Paragraph title: Genome Sequencing ... Incompletely formed SMRTbell templates were digested with a combination of Exonuclease III (New England Biolabs; Ipswich, MA, USA) and Exonuclease VII (Affymetrix; Cleveland, OH, USA).

    Article Title: Genomic mapping of phosphorothioates reveals partial modification of short consensus sequences
    Article Snippet: Paragraph title: Library preparation and SMRT sequencing ... Fragmented DNA was end-repaired, ligated to hairpin adapters and incompletely formed SMRTbell templates were digested with a combination of Exonuclease III (New England Biolabs; Ipswich, MA, USA) and Exonuclease VII (Affymetrix; Cleveland, OH, USA).

    Article Title: Global methylation state at base-pair resolution of the Caulobacter genome throughout the cell cycle
    Article Snippet: Paragraph title: Sequencing. ... Incompletely formed SMRTbell templates were digested with a combination of Exonuclease III (New England Biolabs) and exonuclease VII (Affymetrix).

    Article Title: Human Immunodeficiency Virus Type 1 Nucleocapsid Protein Can Prevent Self-Priming of Minus-Strand Strong Stop DNA by Promoting the Annealing of Short Oligonucleotides to Hairpin Sequences
    Article Snippet: Then 0.3 U of exonuclease VII (Exo VII; Gibco/BRL, Rockville, Md.) was added, and the mixture was incubated for 2 h at 37°C. .. Then 0.3 U of exonuclease VII (Exo VII; Gibco/BRL, Rockville, Md.) was added, and the mixture was incubated for 2 h at 37°C.

    Binding Assay:

    Article Title: Identification of a Pseudomonas aeruginosa PAO1 DNA Methyltransferase, Its Targets, and Physiological Roles
    Article Snippet: DNAs were end repaired and ligated to hairpin adapters by applying components from the DNA/polymerase binding kit 2.0 from Pacific Biosciences, Menlo Park, CA. .. A mixture of exonuclease III and exonuclease VII (Affymetrix, High Wycombe, United Kingdom) was utilized.

    Article Title: First genome sequences of Achromobacter phages reveal new members of the N4 family
    Article Snippet: DNAs were concentrated, end-repaired and ligated to hairpin adapters applying components from the DNA/Polymerase Binding Kit 2.0 from Pacific Biosciences, Menlo Park, CA, USA. .. A mixture of Exonuclease III and Exonuclease VII (Affymetrix, High Wycombe, UK) was utilized.

    DNA Extraction:

    Article Title: A role for the bacterial GATC methylome in antibiotic stress survival
    Article Snippet: Paragraph title: Genomic DNA extraction and PacBio sequencing ... Incompletely formed SMRTbell templates were digested using Exonuclease III (New England Biolabs) and Exonuclease VII (Affymetrix).

    Methylation:

    Article Title: A role for the bacterial GATC methylome in antibiotic stress survival
    Article Snippet: To assess genomic methylation status, gDNA extracted from stationary-phase cultures was quantified, digested using DpnII (NEB) and run on an 0.8% agarose gel containing ethidium bromide. .. Incompletely formed SMRTbell templates were digested using Exonuclease III (New England Biolabs) and Exonuclease VII (Affymetrix).

    Article Title: A random six-phase switch regulates pneumococcal virulence via global epigenetic changes
    Article Snippet: Incompletely formed SMRTbell templates were digested using Exonuclease III (New England Biolabs) and Exonuclease VII (Affymetrix, OH, USA). .. Sequencing was carried out on the PacBio RS II (Menlo Park, CA, USA) using standard protocols for long insert libraries.

    Purification:

    Article Title: First genome sequences of Achromobacter phages reveal new members of the N4 family
    Article Snippet: A mixture of Exonuclease III and Exonuclease VII (Affymetrix, High Wycombe, UK) was utilized. .. A mixture of Exonuclease III and Exonuclease VII (Affymetrix, High Wycombe, UK) was utilized.

    Article Title: Global methylation state at base-pair resolution of the Caulobacter genome throughout the cell cycle
    Article Snippet: Incompletely formed SMRTbell templates were digested with a combination of Exonuclease III (New England Biolabs) and exonuclease VII (Affymetrix). .. Plasmid DNA was fragmented to ∼800 bp, and 5mC residues were converted to 5caC using the 5mC Tet1 Oxidation Kit (WiseGene).

    Article Title: The RecA/RAD51 protein drives migration of Holliday junctions via polymerization on DNA
    Article Snippet: Human RAD51, RAD51K133R , RAD51K133A , and RPA were purified as described ( ). .. SSB, Shrimp alkaline phosphatase, and Exonuclease VII were from USB Corp., and Terminal deoxynucleotidyl transferase, restriction endonucleases, Exonuclease I, and T4 polynucleotide kinase were from New England Biolabs.

    Polymerase Chain Reaction:

    Article Title: A random six-phase switch regulates pneumococcal virulence via global epigenetic changes
    Article Snippet: DNA was extracted from overnight cultures in TSB from each of the different variants using the High Pure PCR Template Preparation kit (Roche, Italy) and sent to Pacific Biosciences (Menlo Park, CA, USA) where methylome data was obtained by SMRT. .. Incompletely formed SMRTbell templates were digested using Exonuclease III (New England Biolabs) and Exonuclease VII (Affymetrix, OH, USA).

    Article Title: Functional Analysis of Three Arabidopsis ARGONAUTES Using Slicer-Defective Mutants
    Article Snippet: Digestion of unligated Cloning Linker 1 was done by first adding 20 units of deadenylase (NEB) to the overnight ligation and incubating at 30°C for 15 min, then adding 10 units of Exonuclease VII (Affymetrix) and incubating at 37°C for 15 min. Next, 15 pmol of RNA 5′ adapter (see online) was denatured at 70°C for 2 min followed by transfer to ice for at least 2 min. Denatured RNA 5′ adapter was added to the ligation mixture with 0.48 mM of ATP and five units of T4 RNA Ligase 1 (Ambion), and the resulting mixture was incubated at 28°C for 1 h, then placed on ice. .. Digestion of unligated Cloning Linker 1 was done by first adding 20 units of deadenylase (NEB) to the overnight ligation and incubating at 30°C for 15 min, then adding 10 units of Exonuclease VII (Affymetrix) and incubating at 37°C for 15 min. Next, 15 pmol of RNA 5′ adapter (see online) was denatured at 70°C for 2 min followed by transfer to ice for at least 2 min. Denatured RNA 5′ adapter was added to the ligation mixture with 0.48 mM of ATP and five units of T4 RNA Ligase 1 (Ambion), and the resulting mixture was incubated at 28°C for 1 h, then placed on ice.

    Immunoprecipitation:

    Article Title: Functional Analysis of Three Arabidopsis ARGONAUTES Using Slicer-Defective Mutants
    Article Snippet: Ten microliters (or 100%) of the immunoprecipitated RNA was mixed with 15 pmol of Cloning Linker 1 (IDT; see online) and incubated at 70°C for 2 min. .. Digestion of unligated Cloning Linker 1 was done by first adding 20 units of deadenylase (NEB) to the overnight ligation and incubating at 30°C for 15 min, then adding 10 units of Exonuclease VII (Affymetrix) and incubating at 37°C for 15 min. Next, 15 pmol of RNA 5′ adapter (see online) was denatured at 70°C for 2 min followed by transfer to ice for at least 2 min. Denatured RNA 5′ adapter was added to the ligation mixture with 0.48 mM of ATP and five units of T4 RNA Ligase 1 (Ambion), and the resulting mixture was incubated at 28°C for 1 h, then placed on ice.

    Agarose Gel Electrophoresis:

    Article Title: A role for the bacterial GATC methylome in antibiotic stress survival
    Article Snippet: To assess genomic methylation status, gDNA extracted from stationary-phase cultures was quantified, digested using DpnII (NEB) and run on an 0.8% agarose gel containing ethidium bromide. .. Incompletely formed SMRTbell templates were digested using Exonuclease III (New England Biolabs) and Exonuclease VII (Affymetrix).

    Plasmid Preparation:

    Article Title: A random six-phase switch regulates pneumococcal virulence via global epigenetic changes
    Article Snippet: Incompletely formed SMRTbell templates were digested using Exonuclease III (New England Biolabs) and Exonuclease VII (Affymetrix, OH, USA). .. Methylation sites of variants SpnIIIA-C were experimentally confirmed by protection of pDP28 DNA from digestion by methylation sensitive enzymes with overlapping target specificity.

    Article Title: Global methylation state at base-pair resolution of the Caulobacter genome throughout the cell cycle
    Article Snippet: Plasmid DNA was sheared to ∼800 bp. .. Incompletely formed SMRTbell templates were digested with a combination of Exonuclease III (New England Biolabs) and exonuclease VII (Affymetrix).

    Software:

    Article Title: DNA‐binding determinants and cellular thresholds for human telomerase repeat addition processivity
    Article Snippet: Reactions were incubated at room temperature for 1 min, followed by addition of five units of ExoVII (Affymetrix). .. Reactions were incubated at room temperature for 1 min, followed by addition of five units of ExoVII (Affymetrix).

    Sample Prep:

    Article Title: First genome sequences of Achromobacter phages reveal new members of the N4 family
    Article Snippet: Paragraph title: Sample preparation and sequencing ... A mixture of Exonuclease III and Exonuclease VII (Affymetrix, High Wycombe, UK) was utilized.

    Concentration Assay:

    Article Title: Human telomerase specialization for repeat synthesis by unique handling of primer-template duplex
    Article Snippet: The primer extension reaction was initiated by the addition, to final concentration, of 50 mM Tris-acetate at pH 8, 3 mM MgCl2 , 1 mM EGTA, 1 mM spermidine, 5 mM DTT, 5% glycerol, 0.1 μM α-32 P dGTP and, if included, 250 μM of other dNTPs and/or 500 μM of ddNTPs in a final volume of 20 μl. .. The reaction mixtures were incubated at room temperature for 1 min, followed by the addition of 5 units of exonuclease VII (ExoVII, Affymetrix).

    Article Title: DNA‐binding determinants and cellular thresholds for human telomerase repeat addition processivity
    Article Snippet: Telomerase immobilized on FLAG‐antibody resin was allowed to bind 5′‐T21 GT2 AG2 ‐3′ primer supplied at 1 μM and incubated at room temperature for 30 min. Primer extension was initiated by addition of reaction buffer and a final concentration of 0.1 μM α‐32 P dGTP and, if included, 250 μM dATP, 250 μM dTTP, and/or 500 μM of ddNTP. .. Reactions were incubated at room temperature for 1 min, followed by addition of five units of ExoVII (Affymetrix).

    Alkaline Lysis:

    Article Title: A random six-phase switch regulates pneumococcal virulence via global epigenetic changes
    Article Snippet: Incompletely formed SMRTbell templates were digested using Exonuclease III (New England Biolabs) and Exonuclease VII (Affymetrix, OH, USA). .. Plasmid DNA for these experiments was extracted from strains expressing a single SpnD39III variant and shown by SMRT sequencing to methylate one single SpnD39III target.

    Variant Assay:

    Article Title: A random six-phase switch regulates pneumococcal virulence via global epigenetic changes
    Article Snippet: Incompletely formed SMRTbell templates were digested using Exonuclease III (New England Biolabs) and Exonuclease VII (Affymetrix, OH, USA). .. Methylation sites of variants SpnIIIA-C were experimentally confirmed by protection of pDP28 DNA from digestion by methylation sensitive enzymes with overlapping target specificity.