q5 dna polymerase  (New England Biolabs)


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    Structured Review

    New England Biolabs q5 dna polymerase
    Q5 Dna Polymerase, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 281 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/q5 dna polymerase/product/New England Biolabs
    Average 99 stars, based on 281 article reviews
    Price from $9.99 to $1999.99
    q5 dna polymerase - by Bioz Stars, 2020-04
    99/100 stars

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    Related Articles

    Clone Assay:

    Article Title: Plasmid Characteristics Modulate the Propensity of Gene Exchange in Bacterial Vesicles
    Article Snippet: All constructs were made via PCR amplification using Q5 DNA polymerase followed by NEB DNA assembly and 42°C heat shock transformation into chemically competent 5-alpha cells (New England BioLabs, Ipswich, MA). .. For pLC-10 and pLC-15 (10 kb and 15 kb, respectively), we started with pLC291 and cloned in lambda DNA from VWR (Radnor, PA) using primers listed in Table S1.

    Amplification:

    Article Title: Plasmid Characteristics Modulate the Propensity of Gene Exchange in Bacterial Vesicles
    Article Snippet: .. All constructs were made via PCR amplification using Q5 DNA polymerase followed by NEB DNA assembly and 42°C heat shock transformation into chemically competent 5-alpha cells (New England BioLabs, Ipswich, MA). .. A 3.5-kb plasmid was constructed from regions of pLC291 that contained the origin of replication and antibiotic resistance from the deposited DNA sequences on Addgene of pLC291; these included three regions: nucleotides (nt) 900 to 1850, 2750 to 5000, and 7200 to 7500.

    Agarose Gel Electrophoresis:

    Article Title: Plasmid Characteristics Modulate the Propensity of Gene Exchange in Bacterial Vesicles
    Article Snippet: All constructs were made via PCR amplification using Q5 DNA polymerase followed by NEB DNA assembly and 42°C heat shock transformation into chemically competent 5-alpha cells (New England BioLabs, Ipswich, MA). .. Confirmation of plasmid size by restriction digestion was performed on 50 ng of purified plasmids with restriction enzyme NotI at 37°C for 1 h and run on a 1% agarose gel (New England BioLabs, Ipswich, MA).

    Synthesized:

    Article Title: A genetic toolbox for metabolic engineering of Issatchenkia orientalis.
    Article Snippet: The nonconventional yeast Issatchenkia orientalis can grow under highly acidic conditions and has been explored for production of various organic acids. .. The nonconventional yeast Issatchenkia orientalis can grow under highly acidic conditions and has been explored for production of various organic acids.

    Mutagenesis:

    Article Title: Plasmid Characteristics Modulate the Propensity of Gene Exchange in Bacterial Vesicles
    Article Snippet: Briefly, using NEB Q5 site-directed mutagenesis, GAG ATT was changed to AAG ATC or CGG ATC, respectively (New England BioLabs, Ipswich, MA). .. All constructs were made via PCR amplification using Q5 DNA polymerase followed by NEB DNA assembly and 42°C heat shock transformation into chemically competent 5-alpha cells (New England BioLabs, Ipswich, MA).

    Isolation:

    Article Title: A genetic toolbox for metabolic engineering of Issatchenkia orientalis.
    Article Snippet: The nonconventional yeast Issatchenkia orientalis can grow under highly acidic conditions and has been explored for production of various organic acids. .. The nonconventional yeast Issatchenkia orientalis can grow under highly acidic conditions and has been explored for production of various organic acids.

    Lambda DNA Preparation:

    Article Title: Plasmid Characteristics Modulate the Propensity of Gene Exchange in Bacterial Vesicles
    Article Snippet: All constructs were made via PCR amplification using Q5 DNA polymerase followed by NEB DNA assembly and 42°C heat shock transformation into chemically competent 5-alpha cells (New England BioLabs, Ipswich, MA). .. For pLC-10 and pLC-15 (10 kb and 15 kb, respectively), we started with pLC291 and cloned in lambda DNA from VWR (Radnor, PA) using primers listed in Table S1.

    Construct:

    Article Title: Plasmid Characteristics Modulate the Propensity of Gene Exchange in Bacterial Vesicles
    Article Snippet: .. All constructs were made via PCR amplification using Q5 DNA polymerase followed by NEB DNA assembly and 42°C heat shock transformation into chemically competent 5-alpha cells (New England BioLabs, Ipswich, MA). .. A 3.5-kb plasmid was constructed from regions of pLC291 that contained the origin of replication and antibiotic resistance from the deposited DNA sequences on Addgene of pLC291; these included three regions: nucleotides (nt) 900 to 1850, 2750 to 5000, and 7200 to 7500.

    Purification:

    Article Title: Plasmid Characteristics Modulate the Propensity of Gene Exchange in Bacterial Vesicles
    Article Snippet: All constructs were made via PCR amplification using Q5 DNA polymerase followed by NEB DNA assembly and 42°C heat shock transformation into chemically competent 5-alpha cells (New England BioLabs, Ipswich, MA). .. Confirmation of plasmid size by restriction digestion was performed on 50 ng of purified plasmids with restriction enzyme NotI at 37°C for 1 h and run on a 1% agarose gel (New England BioLabs, Ipswich, MA).

    SYBR Green Assay:

    Article Title: A genetic toolbox for metabolic engineering of Issatchenkia orientalis.
    Article Snippet: .. The nonconventional yeast Issatchenkia orientalis can grow under highly acidic conditions and has been explored for production of various organic acids. .. The nonconventional yeast Issatchenkia orientalis can grow under highly acidic conditions and has been explored for production of various organic acids.

    other:

    Article Title: A Strongly Fluorescing Anaerobic Reporter and Protein-Tagging System for Clostridium Organisms Based on the Fluorescence-Activating and Absorption-Shifting Tag Protein (FAST)
    Article Snippet: E. coli NEB 5-alpha, Q5 DNA polymerase, and NEBuilder HiFi DNA assembly master mix were purchased from New England BioLabs (NEB).

    Polymerase Chain Reaction:

    Article Title: Plasmid Characteristics Modulate the Propensity of Gene Exchange in Bacterial Vesicles
    Article Snippet: .. All constructs were made via PCR amplification using Q5 DNA polymerase followed by NEB DNA assembly and 42°C heat shock transformation into chemically competent 5-alpha cells (New England BioLabs, Ipswich, MA). .. A 3.5-kb plasmid was constructed from regions of pLC291 that contained the origin of replication and antibiotic resistance from the deposited DNA sequences on Addgene of pLC291; these included three regions: nucleotides (nt) 900 to 1850, 2750 to 5000, and 7200 to 7500.

    Article Title: A genetic toolbox for metabolic engineering of Issatchenkia orientalis.
    Article Snippet: .. The nonconventional yeast Issatchenkia orientalis can grow under highly acidic conditions and has been explored for production of various organic acids. .. The nonconventional yeast Issatchenkia orientalis can grow under highly acidic conditions and has been explored for production of various organic acids.

    Planar Chromatography:

    Article Title: Plasmid Characteristics Modulate the Propensity of Gene Exchange in Bacterial Vesicles
    Article Snippet: All constructs were made via PCR amplification using Q5 DNA polymerase followed by NEB DNA assembly and 42°C heat shock transformation into chemically competent 5-alpha cells (New England BioLabs, Ipswich, MA). .. For pLC-10 and pLC-15 (10 kb and 15 kb, respectively), we started with pLC291 and cloned in lambda DNA from VWR (Radnor, PA) using primers listed in Table S1.

    Transformation Assay:

    Article Title: Plasmid Characteristics Modulate the Propensity of Gene Exchange in Bacterial Vesicles
    Article Snippet: .. All constructs were made via PCR amplification using Q5 DNA polymerase followed by NEB DNA assembly and 42°C heat shock transformation into chemically competent 5-alpha cells (New England BioLabs, Ipswich, MA). .. A 3.5-kb plasmid was constructed from regions of pLC291 that contained the origin of replication and antibiotic resistance from the deposited DNA sequences on Addgene of pLC291; these included three regions: nucleotides (nt) 900 to 1850, 2750 to 5000, and 7200 to 7500.

    Recombinant:

    Article Title: A genetic toolbox for metabolic engineering of Issatchenkia orientalis.
    Article Snippet: The nonconventional yeast Issatchenkia orientalis can grow under highly acidic conditions and has been explored for production of various organic acids. .. The nonconventional yeast Issatchenkia orientalis can grow under highly acidic conditions and has been explored for production of various organic acids.

    Plasmid Preparation:

    Article Title: Plasmid Characteristics Modulate the Propensity of Gene Exchange in Bacterial Vesicles
    Article Snippet: To construct SC101 plasmids with increased plasmid copy numbers, pSC101 was used as the starting plasmid. .. All constructs were made via PCR amplification using Q5 DNA polymerase followed by NEB DNA assembly and 42°C heat shock transformation into chemically competent 5-alpha cells (New England BioLabs, Ipswich, MA).

    Article Title: A genetic toolbox for metabolic engineering of Issatchenkia orientalis.
    Article Snippet: The nonconventional yeast Issatchenkia orientalis can grow under highly acidic conditions and has been explored for production of various organic acids. .. The nonconventional yeast Issatchenkia orientalis can grow under highly acidic conditions and has been explored for production of various organic acids.

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    New England Biolabs q5 dna polymerase
    PCR based verification of genome edit A. A schematic representation showing the locations of PCR primers used in panel B; B. Agarose gel stained with ethidium bromide of amplified PCR products using primers specific to the point mutation (H743A, for which we changed CAC→GCT) and the B. subtilis chromosome (left panel), and using primers specific to the editing plasmid (right panel). Of twelve isolates screened all were found to have the predicted point mutation, and two of twelve retained resistance to spectinomycin (asterisks). The slow migrating band in the PY79 lane corresponds to nonspecific PCR amplification in the wild type background. The editing plasmid was detectable in spectinomycin resistant isolates, but not in the remaining ten. C. Agarose gel stained with ethidium bromide of PCR reactions using primers specific for deletion of four B. subtilis genes. Primers were designed outside of the editing template, and are therefore specific to chromosomal DNA. For all four deletions, all isolates were found to be spectinomycin sensitive, and PCR genotyping of three isolates for each deletion found that all isolates tested had the intended deletion. Note that for the control reaction for ponA , the PCR product is faint because the amplicon is approximately 5 kb and the extension time of the PCR was 3 min, which is near the limit for complete extension for <t>Q5</t> DNA polymerase; the other three control amplicons are no larger than 3.4 kb.
    Q5 Dna Polymerase, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 718 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/q5 dna polymerase/product/New England Biolabs
    Average 99 stars, based on 718 article reviews
    Price from $9.99 to $1999.99
    q5 dna polymerase - by Bioz Stars, 2020-04
    99/100 stars
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    Q5U Hot Start High Fidelity DNA Polymerase is a modified version of Q5 High Fidelity DNA Polymerase which efficiently incorporates dUTP and amplifies uracil containing DNA templates This feature is
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    PCR based verification of genome edit A. A schematic representation showing the locations of PCR primers used in panel B; B. Agarose gel stained with ethidium bromide of amplified PCR products using primers specific to the point mutation (H743A, for which we changed CAC→GCT) and the B. subtilis chromosome (left panel), and using primers specific to the editing plasmid (right panel). Of twelve isolates screened all were found to have the predicted point mutation, and two of twelve retained resistance to spectinomycin (asterisks). The slow migrating band in the PY79 lane corresponds to nonspecific PCR amplification in the wild type background. The editing plasmid was detectable in spectinomycin resistant isolates, but not in the remaining ten. C. Agarose gel stained with ethidium bromide of PCR reactions using primers specific for deletion of four B. subtilis genes. Primers were designed outside of the editing template, and are therefore specific to chromosomal DNA. For all four deletions, all isolates were found to be spectinomycin sensitive, and PCR genotyping of three isolates for each deletion found that all isolates tested had the intended deletion. Note that for the control reaction for ponA , the PCR product is faint because the amplicon is approximately 5 kb and the extension time of the PCR was 3 min, which is near the limit for complete extension for Q5 DNA polymerase; the other three control amplicons are no larger than 3.4 kb.

    Journal: Bio-protocol

    Article Title: CRISPR/Cas9 Editing of the Bacillus subtilis Genome

    doi: 10.21769/BioProtoc.2272

    Figure Lengend Snippet: PCR based verification of genome edit A. A schematic representation showing the locations of PCR primers used in panel B; B. Agarose gel stained with ethidium bromide of amplified PCR products using primers specific to the point mutation (H743A, for which we changed CAC→GCT) and the B. subtilis chromosome (left panel), and using primers specific to the editing plasmid (right panel). Of twelve isolates screened all were found to have the predicted point mutation, and two of twelve retained resistance to spectinomycin (asterisks). The slow migrating band in the PY79 lane corresponds to nonspecific PCR amplification in the wild type background. The editing plasmid was detectable in spectinomycin resistant isolates, but not in the remaining ten. C. Agarose gel stained with ethidium bromide of PCR reactions using primers specific for deletion of four B. subtilis genes. Primers were designed outside of the editing template, and are therefore specific to chromosomal DNA. For all four deletions, all isolates were found to be spectinomycin sensitive, and PCR genotyping of three isolates for each deletion found that all isolates tested had the intended deletion. Note that for the control reaction for ponA , the PCR product is faint because the amplicon is approximately 5 kb and the extension time of the PCR was 3 min, which is near the limit for complete extension for Q5 DNA polymerase; the other three control amplicons are no larger than 3.4 kb.

    Article Snippet: The PCR program used to linearize pPB41 is: PCR amplify CRISPR/Cas9 using plasmid generated in step B1 ( ) Amplify plasmid generated in step B1 (pPB43 in the example) via PCR using Q5 DNA polymerase (NEB) and primers oPEB232 and oPEB234.

    Techniques: Polymerase Chain Reaction, Agarose Gel Electrophoresis, Staining, Amplification, Mutagenesis, Plasmid Preparation

    Assay specificity for PIK 3 CA mutations. Comparison of mutant PIK 3 CA detection in human control genomic DNA ( gDNA ) and cell‐free plasma DNA following pre‐amplification or no pre‐amplification using Q5 High‐Fidelity DNA Polymerase (New England BioLabs) or TaqMan ® PreAmp Master Mix (Life Technologies). The level of blank (LoB) was 0.04% for the mutation E545K and for H1047R, 0.02% for E542K, and less than 0.01% for H1047L as shown by the graphs.

    Journal: Molecular Oncology

    Article Title: Correlation between circulating cell‐free PIK3CA tumor DNA levels and treatment response in patients with PIK3CA‐mutated metastatic breast cancer

    doi: 10.1002/1878-0261.12305

    Figure Lengend Snippet: Assay specificity for PIK 3 CA mutations. Comparison of mutant PIK 3 CA detection in human control genomic DNA ( gDNA ) and cell‐free plasma DNA following pre‐amplification or no pre‐amplification using Q5 High‐Fidelity DNA Polymerase (New England BioLabs) or TaqMan ® PreAmp Master Mix (Life Technologies). The level of blank (LoB) was 0.04% for the mutation E545K and for H1047R, 0.02% for E542K, and less than 0.01% for H1047L as shown by the graphs.

    Article Snippet: To increase the sensitivity of the analysis, the remaining 75 μL serum DNA was subjected to 12 cycles of PCR pre‐amplification with Q5 High‐Fidelity DNA Polymerase (New England BioLabs, Ipswich, MA, USA) using a multiplex PIK3CA primer mix.

    Techniques: Mutagenesis, Amplification

    Deoxynucleoside diphosphate utilization by DNA polymerases. ( A ) A standard PCR with a monophosphate substrate is inhibited, whereas the presence of all four di- or triphosphate nucleotides supports DNA amplification. Thermophilic polymerases Taq, Vent (exo−), and Pfu , as well as B , Deep Vent, and Q5 DNA polymerases, also utilize the diphosphorylated substrates. ( C and D ) Primer-extension reactions on short templates sampled at indicated time points. All reactions were performed with 100 µM dNDPs at 60 °C, except the Bsu reactions, which were performed at 37 °C. Extended sequences are shown alongside of gels and were distinct in C and D . Primer band is indicated with “–P.” Pausing appears mainly before incorporation of dA and dC and is variable among the polymerases. ( E ).

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: DNA synthesis from diphosphate substrates by DNA polymerases

    doi: 10.1073/pnas.1712193115

    Figure Lengend Snippet: Deoxynucleoside diphosphate utilization by DNA polymerases. ( A ) A standard PCR with a monophosphate substrate is inhibited, whereas the presence of all four di- or triphosphate nucleotides supports DNA amplification. Thermophilic polymerases Taq, Vent (exo−), and Pfu , as well as B , Deep Vent, and Q5 DNA polymerases, also utilize the diphosphorylated substrates. ( C and D ) Primer-extension reactions on short templates sampled at indicated time points. All reactions were performed with 100 µM dNDPs at 60 °C, except the Bsu reactions, which were performed at 37 °C. Extended sequences are shown alongside of gels and were distinct in C and D . Primer band is indicated with “–P.” Pausing appears mainly before incorporation of dA and dC and is variable among the polymerases. ( E ).

    Article Snippet: Taq DNA polymerase, Phusion High-Fidelity DNA polymerase, DeepVent DNA polymerase, Vent DNA polymerase, Q5 DNA High-Fidelity polymerase, Bst polymerase, Bsu DNA polymerase (large fragment), and ThermoPol Reaction Buffer [1×: 20 mM Tris⋅HCl, 10 mM (NH4 )2 SO4 , 10 mM KCl, 2 mM MgSO4 , 0.1% Triton–X–100, pH 8.8 at 25 °C] were purchased from New England Biolabs.

    Techniques: Polymerase Chain Reaction, Amplification