10x exonuclease i reaction buffer  (New England Biolabs)


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    Name:
    Exonuclease I Reaction Buffer
    Description:
    Exonuclease I Reaction Buffer 6 0 ml
    Catalog Number:
    b0293s
    Price:
    24
    Size:
    6 0 ml
    Category:
    Buffers
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    Structured Review

    New England Biolabs 10x exonuclease i reaction buffer
    Exonuclease I Reaction Buffer
    Exonuclease I Reaction Buffer 6 0 ml
    https://www.bioz.com/result/10x exonuclease i reaction buffer/product/New England Biolabs
    Average 99 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    10x exonuclease i reaction buffer - by Bioz Stars, 2020-04
    99/100 stars

    Related Products / Commonly Used Together

    exonuclease i enzyme
    exonuclease-treated
    cdna

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    Related Articles

    Transduction:

    Article Title: RemA is a DNA-binding protein that activates biofilm matrix gene expression in Bacillus subtilis
    Article Snippet: The amplified product was transformed into competent cells of PY79 and then transferred to the 3610 background using SPP1-mediated generalized transduction. .. An assembly master mixture was made by combining prepared 5× isothermal assembly reaction buffer (131 mM Tris-HCl, 13.1 mM MgCl2 , 13.1 mM DTT, 8.21 mM PEG-8000, 1.32 mM NAD and 0.26 mM each dNTP) with Phusion DNA polymerase (New England BioLabs) (0.033 units µl−1 ), T5 exonuclease diluted 1:5 with 5× reaction buffer (New England BioLabs) (0.01 units µl−1 ), Taq DNA ligase (New England BioLabs) (5328 units µl−1 ) and additional dNTPs (267 µM).

    Clone Assay:

    Article Title: Light-activated cell identification and sorting (LACIS) for selection of edited clones on a nanofluidic device
    Article Snippet: Sample processing for next-generation sequencing Genomic DNA was extracted from exported clones by incubating in Proteinase K buffer (0.1 M NaCl, 10 mM Tris-HCl pH 8.0, 1 mM EDTA) for 30 min at 55 °C, then for 20 min at 80 °C to inactivate Proteinase K. The genomic region around the CRISPR/Cas9 target site for CXCR4 gene was amplified by PCR with primers positioned outside of the HDR repair template sequence (positioned to avoid amplification of exogenous template) for ten cycles using KAPA HiFi Hotstart ReadyMix (Kapa Biosystems, KR0370) according to the manufacturer’s protocol (PCR primers listed in Supplementary Data ). .. Excess PCR primers were removed by incubating with Exonuclease I (NEB, M0293S) in 1 × exonuclease reaction buffer (NEB, B0293S) for 1 h at 37 °C, followed by enzyme inactivation for 20 min at 80 °C.

    Amplification:

    Article Title: Light-activated cell identification and sorting (LACIS) for selection of edited clones on a nanofluidic device
    Article Snippet: Sample processing for next-generation sequencing Genomic DNA was extracted from exported clones by incubating in Proteinase K buffer (0.1 M NaCl, 10 mM Tris-HCl pH 8.0, 1 mM EDTA) for 30 min at 55 °C, then for 20 min at 80 °C to inactivate Proteinase K. The genomic region around the CRISPR/Cas9 target site for CXCR4 gene was amplified by PCR with primers positioned outside of the HDR repair template sequence (positioned to avoid amplification of exogenous template) for ten cycles using KAPA HiFi Hotstart ReadyMix (Kapa Biosystems, KR0370) according to the manufacturer’s protocol (PCR primers listed in Supplementary Data ). .. Excess PCR primers were removed by incubating with Exonuclease I (NEB, M0293S) in 1 × exonuclease reaction buffer (NEB, B0293S) for 1 h at 37 °C, followed by enzyme inactivation for 20 min at 80 °C.

    Article Title: RemA is a DNA-binding protein that activates biofilm matrix gene expression in Bacillus subtilis
    Article Snippet: Insertions were verified by PCR amplification using primers 3318/3321. .. An assembly master mixture was made by combining prepared 5× isothermal assembly reaction buffer (131 mM Tris-HCl, 13.1 mM MgCl2 , 13.1 mM DTT, 8.21 mM PEG-8000, 1.32 mM NAD and 0.26 mM each dNTP) with Phusion DNA polymerase (New England BioLabs) (0.033 units µl−1 ), T5 exonuclease diluted 1:5 with 5× reaction buffer (New England BioLabs) (0.01 units µl−1 ), Taq DNA ligase (New England BioLabs) (5328 units µl−1 ) and additional dNTPs (267 µM).

    Article Title: Spatial distribution and function of T follicular regulatory cells in human lymph nodes
    Article Snippet: Then, each sample was reverse-transcribed using Reverse Transcription Mastermix (Fluidigm). cDNA was amplified with 15 cycles of specific target preamplification using the Fluidigm Pre-Amp Master Mix and the full complement of primers. .. Preamplified samples were subjected to an exonuclease reaction, using Exonuclease I and Exonuclease I Reaction Buffer (New England Biolabs).

    Article Title: Decontamination of 16S rRNA gene amplicon sequence datasets based on bacterial load assessment by qPCR
    Article Snippet: PCR and sequencing The V3–4 region of the bacterial 16S rRNA genes (E. coli positions 341–805) was amplified using template DNA from E. coli and S. aureus cultures, and from NECs. .. The primers from the first round of PCR were removed by digesting 5-μl samples with 1 unit Exonuclease I (New England Biolabs, Ipswich, MA, USA) in a total volume of 10 μl Exonuclease I Reaction Buffer (New England Biolabs) at 37 °C for 30 min.

    Article Title: Non-coding antisense transcription detected by conventional and single-stranded cDNA microarray
    Article Snippet: The test slides also included background control spots produced by processing PCR reactions without template (PCR primers with no amplification product). .. Slides were overlaid with 45 μl exonuclease reaction mixture (1 U/μl T7 exonuclease 6 in 1× reaction buffer, New England Biolabs), covered with a coverslip and left to incubate for 30 minutes at 25°C in a CMT Corning hybridization chamber.

    Article Title: Endovascular biopsy: Strategy for analyzing gene expression profiles of individual endothelial cells obtained from human vessels✩
    Article Snippet: The samples were incubated at 50 °C for 15 min for the reverse transcription, 95 °C for 2 min for inactivating reverse transcriptase and activating Taq polymerase, then subjected to 18 PCR cycles (95 °C 15 sec then 60 °C for 4 min for each cycle) for specific targets amplification (STA). .. To remove the unincorporated primers for best results, each sample was then mixed with 3.6 μL exonuclease treatment buffer composed of 2.52 μL water, 0.36 μL 10× Exonuclease I reaction buffer and 0.72 μL 20 units/μL Exonuclease I (New England BioLabs, Ipswich, MA), incubated at 37 °C for 30 min for digestion and 80 °C for 15 min to inactivate the exonuclease.

    Quantitative RT-PCR:

    Article Title: Spatial distribution and function of T follicular regulatory cells in human lymph nodes
    Article Snippet: Paragraph title: Real-time quantitative RT-PCR ... Preamplified samples were subjected to an exonuclease reaction, using Exonuclease I and Exonuclease I Reaction Buffer (New England Biolabs).

    Incubation:

    Article Title: Simplified ChIP-exo assays
    Article Snippet: .. The λ exonuclease digestion (100 µl) containing: 20 U λ exonuclease (NEB), 1 × λ exonuclease reaction buffer (NEB), 0.1% Triton-X 100, and 5% DMSO was incubated for 30 min at 37 °C; then washed with 10 mM Tris-HCl, pH 8.0 at 4 °C. .. The RecJf exonuclease digestion (100 µl) containing: 75 U RecJf exonuclease (NEB), 2 × NEBuffer 2, 0.1% Triton-X 100, and 5% DMSO was incubated for 30 min at 37 °C; then washed with 10 mM Tris-HCl, pH 8.0 at 4 °C.

    Article Title: Endovascular biopsy: Strategy for analyzing gene expression profiles of individual endothelial cells obtained from human vessels✩
    Article Snippet: .. To remove the unincorporated primers for best results, each sample was then mixed with 3.6 μL exonuclease treatment buffer composed of 2.52 μL water, 0.36 μL 10× Exonuclease I reaction buffer and 0.72 μL 20 units/μL Exonuclease I (New England BioLabs, Ipswich, MA), incubated at 37 °C for 30 min for digestion and 80 °C for 15 min to inactivate the exonuclease. .. Quantitative RT-PCR 48.48 nanofluidic chips and a BioMark HD system (Fluidigm, South San Francisco, CA) were used.

    Expressing:

    Article Title: Limited immune surveillance in lymphoid tissue by cytolytic CD4+ T cells during health and HIV disease
    Article Snippet: Paragraph title: Single-cell gene expression analysis ... Exonuclease mixture, containing of Exonuclease I (New England BioLabs), 10× Exonuclease I Reaction Buffer (New England BioLabs) and H2 O was added to each well to remove excessive primers and the plate was run on a thermocycler (37°C for 30 min, 80°C for 15 min).

    Transformation Assay:

    Article Title: RemA is a DNA-binding protein that activates biofilm matrix gene expression in Bacillus subtilis
    Article Snippet: The amplified product was transformed into competent cells of PY79 and then transferred to the 3610 background using SPP1-mediated generalized transduction. .. An assembly master mixture was made by combining prepared 5× isothermal assembly reaction buffer (131 mM Tris-HCl, 13.1 mM MgCl2 , 13.1 mM DTT, 8.21 mM PEG-8000, 1.32 mM NAD and 0.26 mM each dNTP) with Phusion DNA polymerase (New England BioLabs) (0.033 units µl−1 ), T5 exonuclease diluted 1:5 with 5× reaction buffer (New England BioLabs) (0.01 units µl−1 ), Taq DNA ligase (New England BioLabs) (5328 units µl−1 ) and additional dNTPs (267 µM).

    Hybridization:

    Article Title: Non-coding antisense transcription detected by conventional and single-stranded cDNA microarray
    Article Snippet: .. Slides were overlaid with 45 μl exonuclease reaction mixture (1 U/μl T7 exonuclease 6 in 1× reaction buffer, New England Biolabs), covered with a coverslip and left to incubate for 30 minutes at 25°C in a CMT Corning hybridization chamber. .. Following in situ digestion, probes were denatured by immersing the arrays in boiling water for two minutes.

    Generated:

    Article Title: Limited immune surveillance in lymphoid tissue by cytolytic CD4+ T cells during health and HIV disease
    Article Snippet: Exonuclease mixture, containing of Exonuclease I (New England BioLabs), 10× Exonuclease I Reaction Buffer (New England BioLabs) and H2 O was added to each well to remove excessive primers and the plate was run on a thermocycler (37°C for 30 min, 80°C for 15 min). .. Distinct primer assays were generated by adding individual primer pairs (5μM) together with a mix of 2x Assay Loading Reagent (Fluidigm) and 1x DNA suspension buffer to each well of a new plate.

    other:

    Article Title: Transcriptional Regulation and Mechanism of SigN (ZpdN), a pBS32-Encoded Sigma Factor in Bacillus subtilis
    Article Snippet: An assembly master mixture was made by combining prepared 5× isothermal assembly reaction buffer (131 mM Tris-HCl, 13.1 mM MgCl2 , 13.1 mM DTT, 8.21 mM PEG 8000, 1.32 mM NAD, and 0.26 mM [each] dNTP) with Phusion DNA polymerase (New England BioLabs) (0.033 units/μl), T5 exonuclease diluted 1:5 with 5× reaction buffer (New England BioLabs) (0.01 units/μl), Taq DNA ligase (New England BioLabs) (5,328 units/μl), and additional dNTPs (267 μM).

    Polymerase Chain Reaction:

    Article Title: Light-activated cell identification and sorting (LACIS) for selection of edited clones on a nanofluidic device
    Article Snippet: .. Excess PCR primers were removed by incubating with Exonuclease I (NEB, M0293S) in 1 × exonuclease reaction buffer (NEB, B0293S) for 1 h at 37 °C, followed by enzyme inactivation for 20 min at 80 °C. .. Amplicon pools were re-amplified by PCR for 15 cycles using a universal primer to add the sequencing adaptor and secondary barcodes to allow parallel sequencing of multiple amplicon pools (Primer sequences are listed in Supplementary Data ).

    Article Title: Limited immune surveillance in lymphoid tissue by cytolytic CD4+ T cells during health and HIV disease
    Article Snippet: Similar to before [ ], PCR plates were thawed and pre-heated for 90 seconds at 65°C. .. Exonuclease mixture, containing of Exonuclease I (New England BioLabs), 10× Exonuclease I Reaction Buffer (New England BioLabs) and H2 O was added to each well to remove excessive primers and the plate was run on a thermocycler (37°C for 30 min, 80°C for 15 min).

    Article Title: RemA is a DNA-binding protein that activates biofilm matrix gene expression in Bacillus subtilis
    Article Snippet: Insertions were verified by PCR amplification using primers 3318/3321. .. An assembly master mixture was made by combining prepared 5× isothermal assembly reaction buffer (131 mM Tris-HCl, 13.1 mM MgCl2 , 13.1 mM DTT, 8.21 mM PEG-8000, 1.32 mM NAD and 0.26 mM each dNTP) with Phusion DNA polymerase (New England BioLabs) (0.033 units µl−1 ), T5 exonuclease diluted 1:5 with 5× reaction buffer (New England BioLabs) (0.01 units µl−1 ), Taq DNA ligase (New England BioLabs) (5328 units µl−1 ) and additional dNTPs (267 µM).

    Article Title: Decontamination of 16S rRNA gene amplicon sequence datasets based on bacterial load assessment by qPCR
    Article Snippet: .. The primers from the first round of PCR were removed by digesting 5-μl samples with 1 unit Exonuclease I (New England Biolabs, Ipswich, MA, USA) in a total volume of 10 μl Exonuclease I Reaction Buffer (New England Biolabs) at 37 °C for 30 min. .. The enzyme was inactivated at 95 °C for 15 min. Amplicon barcoding was performed by re-amplification using 1 μl of Exonuclease I-treated first-round PCR, 15 pmol each of forward primer 5’-NNNNNNNNNNTCCTACGGGNGGCWGCAG-3’ and reverse primer 5’- NNNNNNNNNNTGACTACHVGGGTATCTAAKCC-3’ in a 20-μL volume of MyTaq buffer that contained 1.5 units MyTaq DNA polymerase (Bioline, London, UK) and 2 μl of BioStab PCR optimizer (II) (Sigma-Aldrich, Munich, Germany).

    Article Title: ChIP-seq: using high-throughput sequencing to discover protein-DNA interactions
    Article Snippet: .. Exonuclease digestion of linkers on phosphorylated LMPCR library: Mix directly on ice in a PCR tube: 8.5 μl sample; 1 μl 10 × reaction buffer (NEB, B0262S) and 0.5 μl Lambda exonuclease (NEB, M0262S). .. Digest for 10 min at 37 °C in a PCR block.

    Article Title: Non-coding antisense transcription detected by conventional and single-stranded cDNA microarray
    Article Snippet: The test slides also included background control spots produced by processing PCR reactions without template (PCR primers with no amplification product). .. Slides were overlaid with 45 μl exonuclease reaction mixture (1 U/μl T7 exonuclease 6 in 1× reaction buffer, New England Biolabs), covered with a coverslip and left to incubate for 30 minutes at 25°C in a CMT Corning hybridization chamber.

    Article Title: Endovascular biopsy: Strategy for analyzing gene expression profiles of individual endothelial cells obtained from human vessels✩
    Article Snippet: The samples were incubated at 50 °C for 15 min for the reverse transcription, 95 °C for 2 min for inactivating reverse transcriptase and activating Taq polymerase, then subjected to 18 PCR cycles (95 °C 15 sec then 60 °C for 4 min for each cycle) for specific targets amplification (STA). .. To remove the unincorporated primers for best results, each sample was then mixed with 3.6 μL exonuclease treatment buffer composed of 2.52 μL water, 0.36 μL 10× Exonuclease I reaction buffer and 0.72 μL 20 units/μL Exonuclease I (New England BioLabs, Ipswich, MA), incubated at 37 °C for 30 min for digestion and 80 °C for 15 min to inactivate the exonuclease.

    Binding Assay:

    Article Title: Limited immune surveillance in lymphoid tissue by cytolytic CD4+ T cells during health and HIV disease
    Article Snippet: Exonuclease mixture, containing of Exonuclease I (New England BioLabs), 10× Exonuclease I Reaction Buffer (New England BioLabs) and H2 O was added to each well to remove excessive primers and the plate was run on a thermocycler (37°C for 30 min, 80°C for 15 min). .. A “sample PCR plate” was created by dispensing a sample master mix, containing 2x Sso Fast EvaGreen Supermix with Low ROX (Bio-Rad), 20x DNA Binding Dye Sample Loading Reagent (Fluidigm) and H2O to each well.

    Article Title: Spatial distribution and function of T follicular regulatory cells in human lymph nodes
    Article Snippet: Preamplified samples were subjected to an exonuclease reaction, using Exonuclease I and Exonuclease I Reaction Buffer (New England Biolabs). .. An aliquot of each of the diluted, preamplified samples was then mixed with SsoFast EvaGreen Supermix with Low Rox (Bio-Rad) and DNA Binding Dye Sample Loading Reagent (Fluidigm).

    ChIP-chip:

    Article Title: ChIP-seq: using high-throughput sequencing to discover protein-DNA interactions
    Article Snippet: Paragraph title: 2.2.13. Optional rescue of traditional ChIP-chip libraries for ChIP-seq ... Exonuclease digestion of linkers on phosphorylated LMPCR library: Mix directly on ice in a PCR tube: 8.5 μl sample; 1 μl 10 × reaction buffer (NEB, B0262S) and 0.5 μl Lambda exonuclease (NEB, M0262S).

    ChIP-sequencing:

    Article Title: ChIP-seq: using high-throughput sequencing to discover protein-DNA interactions
    Article Snippet: Paragraph title: 2.2.13. Optional rescue of traditional ChIP-chip libraries for ChIP-seq ... Exonuclease digestion of linkers on phosphorylated LMPCR library: Mix directly on ice in a PCR tube: 8.5 μl sample; 1 μl 10 × reaction buffer (NEB, B0262S) and 0.5 μl Lambda exonuclease (NEB, M0262S).

    Nucleic Acid Electrophoresis:

    Article Title: Decontamination of 16S rRNA gene amplicon sequence datasets based on bacterial load assessment by qPCR
    Article Snippet: The primers from the first round of PCR were removed by digesting 5-μl samples with 1 unit Exonuclease I (New England Biolabs, Ipswich, MA, USA) in a total volume of 10 μl Exonuclease I Reaction Buffer (New England Biolabs) at 37 °C for 30 min. .. PCRs were carried out using the following parameters: pre-denaturation for 2 min at 96 °C, followed by eight cycles of 96 °C for 15 s, 50 °C for 30 s, and 70 °C for 90 s. DNA concentration of amplicons of interest was determined by gel electrophoresis.

    Magnetic Beads:

    Article Title: Simplified ChIP-exo assays
    Article Snippet: The λ exonuclease digestion (100 µl) containing: 20 U λ exonuclease (NEB), 1 × λ exonuclease reaction buffer (NEB), 0.1% Triton-X 100, and 5% DMSO was incubated for 30 min at 37 °C; then washed with 10 mM Tris-HCl, pH 8.0 at 4 °C. .. The supernatant was then transferred to a new tube and purified with Agencourt AMPure magnetic beads (Beckman Coulter) following manufacturer's instructions and using 1.8 × volume of AMPure slurry added to the DNA volume (72 µl).The sample was eluted from the AMPure beads in 10 µl of water, and the following enzymatic steps were carried out in solution.

    Isolation:

    Article Title: Light-activated cell identification and sorting (LACIS) for selection of edited clones on a nanofluidic device
    Article Snippet: Excess PCR primers were removed by incubating with Exonuclease I (NEB, M0293S) in 1 × exonuclease reaction buffer (NEB, B0293S) for 1 h at 37 °C, followed by enzyme inactivation for 20 min at 80 °C. .. PCR products of the expected size were isolated with Select-A-Size DNA Clean and Concentrator (Zymo research, D4080) as sequencing libraries.

    Article Title: Spatial distribution and function of T follicular regulatory cells in human lymph nodes
    Article Snippet: RNA was first isolated from sorted cell populations using the RNeasy Micro Plus kit (Qiagen). .. Preamplified samples were subjected to an exonuclease reaction, using Exonuclease I and Exonuclease I Reaction Buffer (New England Biolabs).

    Size-exclusion Chromatography:

    Article Title: Limited immune surveillance in lymphoid tissue by cytolytic CD4+ T cells during health and HIV disease
    Article Snippet: Next, pre-amplification mix, consisting of pooled mixture of all primer assays (500nM), 5× PreAmp Master Mix (Fluidigm) and H2 O, was added to each well and run on a thermocycler (95°C for 5 min followed by 18 cycles: 96°C for 5 sec 60°C for 6 min). .. Exonuclease mixture, containing of Exonuclease I (New England BioLabs), 10× Exonuclease I Reaction Buffer (New England BioLabs) and H2 O was added to each well to remove excessive primers and the plate was run on a thermocycler (37°C for 30 min, 80°C for 15 min).

    Article Title: Endovascular biopsy: Strategy for analyzing gene expression profiles of individual endothelial cells obtained from human vessels✩
    Article Snippet: The samples were incubated at 50 °C for 15 min for the reverse transcription, 95 °C for 2 min for inactivating reverse transcriptase and activating Taq polymerase, then subjected to 18 PCR cycles (95 °C 15 sec then 60 °C for 4 min for each cycle) for specific targets amplification (STA). .. To remove the unincorporated primers for best results, each sample was then mixed with 3.6 μL exonuclease treatment buffer composed of 2.52 μL water, 0.36 μL 10× Exonuclease I reaction buffer and 0.72 μL 20 units/μL Exonuclease I (New England BioLabs, Ipswich, MA), incubated at 37 °C for 30 min for digestion and 80 °C for 15 min to inactivate the exonuclease.

    Purification:

    Article Title: Simplified ChIP-exo assays
    Article Snippet: The λ exonuclease digestion (100 µl) containing: 20 U λ exonuclease (NEB), 1 × λ exonuclease reaction buffer (NEB), 0.1% Triton-X 100, and 5% DMSO was incubated for 30 min at 37 °C; then washed with 10 mM Tris-HCl, pH 8.0 at 4 °C. .. The supernatant was then transferred to a new tube and purified with Agencourt AMPure magnetic beads (Beckman Coulter) following manufacturer's instructions and using 1.8 × volume of AMPure slurry added to the DNA volume (72 µl).The sample was eluted from the AMPure beads in 10 µl of water, and the following enzymatic steps were carried out in solution.

    Sequencing:

    Article Title: Light-activated cell identification and sorting (LACIS) for selection of edited clones on a nanofluidic device
    Article Snippet: Sample processing for next-generation sequencing Genomic DNA was extracted from exported clones by incubating in Proteinase K buffer (0.1 M NaCl, 10 mM Tris-HCl pH 8.0, 1 mM EDTA) for 30 min at 55 °C, then for 20 min at 80 °C to inactivate Proteinase K. The genomic region around the CRISPR/Cas9 target site for CXCR4 gene was amplified by PCR with primers positioned outside of the HDR repair template sequence (positioned to avoid amplification of exogenous template) for ten cycles using KAPA HiFi Hotstart ReadyMix (Kapa Biosystems, KR0370) according to the manufacturer’s protocol (PCR primers listed in Supplementary Data ). .. Excess PCR primers were removed by incubating with Exonuclease I (NEB, M0293S) in 1 × exonuclease reaction buffer (NEB, B0293S) for 1 h at 37 °C, followed by enzyme inactivation for 20 min at 80 °C.

    Article Title: Decontamination of 16S rRNA gene amplicon sequence datasets based on bacterial load assessment by qPCR
    Article Snippet: Paragraph title: PCR and sequencing ... The primers from the first round of PCR were removed by digesting 5-μl samples with 1 unit Exonuclease I (New England Biolabs, Ipswich, MA, USA) in a total volume of 10 μl Exonuclease I Reaction Buffer (New England Biolabs) at 37 °C for 30 min.

    FACS:

    Article Title: Limited immune surveillance in lymphoid tissue by cytolytic CD4+ T cells during health and HIV disease
    Article Snippet: After FACS sorting, PCR plates were frozen and kept in -80°C until usage. .. Exonuclease mixture, containing of Exonuclease I (New England BioLabs), 10× Exonuclease I Reaction Buffer (New England BioLabs) and H2 O was added to each well to remove excessive primers and the plate was run on a thermocycler (37°C for 30 min, 80°C for 15 min).

    Blocking Assay:

    Article Title: ChIP-seq: using high-throughput sequencing to discover protein-DNA interactions
    Article Snippet: Exonuclease digestion of linkers on phosphorylated LMPCR library: Mix directly on ice in a PCR tube: 8.5 μl sample; 1 μl 10 × reaction buffer (NEB, B0262S) and 0.5 μl Lambda exonuclease (NEB, M0262S). .. Digest for 10 min at 37 °C in a PCR block.

    CRISPR:

    Article Title: Light-activated cell identification and sorting (LACIS) for selection of edited clones on a nanofluidic device
    Article Snippet: Sample processing for next-generation sequencing Genomic DNA was extracted from exported clones by incubating in Proteinase K buffer (0.1 M NaCl, 10 mM Tris-HCl pH 8.0, 1 mM EDTA) for 30 min at 55 °C, then for 20 min at 80 °C to inactivate Proteinase K. The genomic region around the CRISPR/Cas9 target site for CXCR4 gene was amplified by PCR with primers positioned outside of the HDR repair template sequence (positioned to avoid amplification of exogenous template) for ten cycles using KAPA HiFi Hotstart ReadyMix (Kapa Biosystems, KR0370) according to the manufacturer’s protocol (PCR primers listed in Supplementary Data ). .. Excess PCR primers were removed by incubating with Exonuclease I (NEB, M0293S) in 1 × exonuclease reaction buffer (NEB, B0293S) for 1 h at 37 °C, followed by enzyme inactivation for 20 min at 80 °C.

    Chromatin Immunoprecipitation:

    Article Title: Simplified ChIP-exo assays
    Article Snippet: Paragraph title: ChIP-exo 3.0 and 3.1 (tagmentation-based version) ... The λ exonuclease digestion (100 µl) containing: 20 U λ exonuclease (NEB), 1 × λ exonuclease reaction buffer (NEB), 0.1% Triton-X 100, and 5% DMSO was incubated for 30 min at 37 °C; then washed with 10 mM Tris-HCl, pH 8.0 at 4 °C.

    In Situ:

    Article Title: Non-coding antisense transcription detected by conventional and single-stranded cDNA microarray
    Article Snippet: The deposited dsDNA was subsequently digested in situ with T7 exonucelase 6. .. Slides were overlaid with 45 μl exonuclease reaction mixture (1 U/μl T7 exonuclease 6 in 1× reaction buffer, New England Biolabs), covered with a coverslip and left to incubate for 30 minutes at 25°C in a CMT Corning hybridization chamber.

    Next-Generation Sequencing:

    Article Title: Light-activated cell identification and sorting (LACIS) for selection of edited clones on a nanofluidic device
    Article Snippet: Paragraph title: Sample processing for next-generation sequencing ... Excess PCR primers were removed by incubating with Exonuclease I (NEB, M0293S) in 1 × exonuclease reaction buffer (NEB, B0293S) for 1 h at 37 °C, followed by enzyme inactivation for 20 min at 80 °C.

    Produced:

    Article Title: Non-coding antisense transcription detected by conventional and single-stranded cDNA microarray
    Article Snippet: The test slides also included background control spots produced by processing PCR reactions without template (PCR primers with no amplification product). .. Slides were overlaid with 45 μl exonuclease reaction mixture (1 U/μl T7 exonuclease 6 in 1× reaction buffer, New England Biolabs), covered with a coverslip and left to incubate for 30 minutes at 25°C in a CMT Corning hybridization chamber.

    Concentration Assay:

    Article Title: Spatial distribution and function of T follicular regulatory cells in human lymph nodes
    Article Snippet: Preamplified samples were subjected to an exonuclease reaction, using Exonuclease I and Exonuclease I Reaction Buffer (New England Biolabs). .. Assays were prepared using 100-µM primers combined with 2× Assay Loading Reagent (Fluidigm) and DNA suspension buffer (TEKnova), for a final primer concentration of 5 µM.

    Article Title: Decontamination of 16S rRNA gene amplicon sequence datasets based on bacterial load assessment by qPCR
    Article Snippet: The primers from the first round of PCR were removed by digesting 5-μl samples with 1 unit Exonuclease I (New England Biolabs, Ipswich, MA, USA) in a total volume of 10 μl Exonuclease I Reaction Buffer (New England Biolabs) at 37 °C for 30 min. .. PCRs were carried out using the following parameters: pre-denaturation for 2 min at 96 °C, followed by eight cycles of 96 °C for 15 s, 50 °C for 30 s, and 70 °C for 90 s. DNA concentration of amplicons of interest was determined by gel electrophoresis.

    Article Title: Simplified ChIP-exo assays
    Article Snippet: The tagmentation reaction (30 µl) containing: 20 mM Tris-HCl, pH 7.5, 5 mM MgCl2, 10% dimethylformamide, and 1 × Tagmentation Mix from Step 1 (final concentration: 1.25 µM Tn5, 5% glycerol, 750 nM adapter) was incubated for 30 min at 37 °C. .. The λ exonuclease digestion (100 µl) containing: 20 U λ exonuclease (NEB), 1 × λ exonuclease reaction buffer (NEB), 0.1% Triton-X 100, and 5% DMSO was incubated for 30 min at 37 °C; then washed with 10 mM Tris-HCl, pH 8.0 at 4 °C.

    Lysis:

    Article Title: Simplified ChIP-exo assays
    Article Snippet: The ChIP material on resin was washed sequentially with FA Lysis Buffer, NaCl Buffer (50 mM HEPES-KOH, pH 7.5, 500 mM NaCl, 2 mM EDTA, 1% Triton-X 100, 0.1% sodium deoxycholate), LiCl Buffer (100 mM Tris-HCl, pH 8.0, 500 mM LiCl, 1% NP-40, 1% sodium deoxycholate), and 10 mM Tris-HCl, pH 8.0 at 4 °C. .. The λ exonuclease digestion (100 µl) containing: 20 U λ exonuclease (NEB), 1 × λ exonuclease reaction buffer (NEB), 0.1% Triton-X 100, and 5% DMSO was incubated for 30 min at 37 °C; then washed with 10 mM Tris-HCl, pH 8.0 at 4 °C.

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    New England Biolabs 10x exonuclease i reaction buffer
    10x Exonuclease I Reaction Buffer, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/10x exonuclease i reaction buffer/product/New England Biolabs
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