klenow dna polymerase  (New England Biolabs)


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  • 99
    Name:
    DNA Polymerase I Klenow Fragment
    Description:
    DNA Polymerase I Klenow Fragment 1 000 units
    Catalog Number:
    m0210l
    Price:
    248
    Size:
    1 000 units
    Category:
    DNA Polymerases
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    Structured Review

    New England Biolabs klenow dna polymerase
    DNA Polymerase I Klenow Fragment
    DNA Polymerase I Klenow Fragment 1 000 units
    https://www.bioz.com/result/klenow dna polymerase/product/New England Biolabs
    Average 99 stars, based on 26 article reviews
    Price from $9.99 to $1999.99
    klenow dna polymerase - by Bioz Stars, 2020-07
    99/100 stars

    Images

    1) Product Images from "An Efficient Strategy for Broad-Range Detection of Low Abundance Bacteria without DNA Decontamination of PCR Reagents"

    Article Title: An Efficient Strategy for Broad-Range Detection of Low Abundance Bacteria without DNA Decontamination of PCR Reagents

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0020303

    The principle of PE-PCR for bacterial DNA amplification and detection. A fusion probe is designed with the sequences at the 3′-end corresponding to the bacterial genomic sequences and a non-bacterial tag sequence at the 5′-end. The reaction is initiated by annealing the fusion probe to the template bacterial DNA after heat-denaturing at 95°C for 5 min (Step 1 and 2). An enzyme mix (EK mix) of exo I and Klenow DNA polymerase is then added into the reaction mixture and incubated at 37°C for 2 h (Step 3a and 3b). Following heat-inactivation of EK mix at 80°C for 20 min (Step 3c), a forward primer (non-bac-F) corresponding to the non-bacterial sequence of the fusion probe and a reverse primer (bac-R) targeting bacterial genomic sequence downstream of the fusion probe are used for PCR amplification of the primer extension product (Step 4). In this setting, only template bacterial DNA but not the endogenous contaminated bacterial DNA is amplified (Step 5).
    Figure Legend Snippet: The principle of PE-PCR for bacterial DNA amplification and detection. A fusion probe is designed with the sequences at the 3′-end corresponding to the bacterial genomic sequences and a non-bacterial tag sequence at the 5′-end. The reaction is initiated by annealing the fusion probe to the template bacterial DNA after heat-denaturing at 95°C for 5 min (Step 1 and 2). An enzyme mix (EK mix) of exo I and Klenow DNA polymerase is then added into the reaction mixture and incubated at 37°C for 2 h (Step 3a and 3b). Following heat-inactivation of EK mix at 80°C for 20 min (Step 3c), a forward primer (non-bac-F) corresponding to the non-bacterial sequence of the fusion probe and a reverse primer (bac-R) targeting bacterial genomic sequence downstream of the fusion probe are used for PCR amplification of the primer extension product (Step 4). In this setting, only template bacterial DNA but not the endogenous contaminated bacterial DNA is amplified (Step 5).

    Techniques Used: Polymerase Chain Reaction, Amplification, Genomic Sequencing, Sequencing, Incubation, BAC Assay

    2) Product Images from "Strand displacement synthesis by yeast DNA polymerase ε"

    Article Title: Strand displacement synthesis by yeast DNA polymerase ε

    Journal: Nucleic Acids Research

    doi: 10.1093/nar/gkw556

    Pol ε poorly extends D-loops in comparison to Pol δ but is proficient to extend primed single-stranded DNA using the same substrates under the same conditions. ( A ) In vitro D-loop reactions using a 37-mer oligonucleotide were reconstituted using purified S. cerevisiae proteins as described in Materials and Methods. ( B ) Product analysis of reconstituted D-loop reactions containing either Klenow polymerase, Pol δ (10 nM) or Pol ε (10 nM) at 0, 2 (not for Klenow), 5 and 10 min extension times. ( C ) Extension of primed single-stranded circular template DNA using a 37-mer oligonucleotide. ( D ) Product analysis of primer extension on denaturing gels of reaction containing Klenow polymerase, Pol δ (10 nM) or Pol ε (10 nM) each plus or minus 10 nM PCNA/RFC at 0, 2 (not for Klenow), 5 and 10 min extension times. A 100 nt size ladder is shown in the left-most lane.
    Figure Legend Snippet: Pol ε poorly extends D-loops in comparison to Pol δ but is proficient to extend primed single-stranded DNA using the same substrates under the same conditions. ( A ) In vitro D-loop reactions using a 37-mer oligonucleotide were reconstituted using purified S. cerevisiae proteins as described in Materials and Methods. ( B ) Product analysis of reconstituted D-loop reactions containing either Klenow polymerase, Pol δ (10 nM) or Pol ε (10 nM) at 0, 2 (not for Klenow), 5 and 10 min extension times. ( C ) Extension of primed single-stranded circular template DNA using a 37-mer oligonucleotide. ( D ) Product analysis of primer extension on denaturing gels of reaction containing Klenow polymerase, Pol δ (10 nM) or Pol ε (10 nM) each plus or minus 10 nM PCNA/RFC at 0, 2 (not for Klenow), 5 and 10 min extension times. A 100 nt size ladder is shown in the left-most lane.

    Techniques Used: In Vitro, Purification

    3) Product Images from "Zinc-finger nuclease-driven targeted integration into mammalian genomes using donors with limited chromosomal homology"

    Article Title: Zinc-finger nuclease-driven targeted integration into mammalian genomes using donors with limited chromosomal homology

    Journal: Nucleic Acids Research

    doi: 10.1093/nar/gkq512

    Analysis of the overhang types created by ZFNs. ( A ) Scheme to determine ZFN overhangs. A supercoiled plasmid with a ZFN cleavage site is cut by a titration of in vitro transcribed and translated ZFNs. ZFN-linearized plasmids are purified by gel electrophoresis, 5′ overhangs filled in with Klenow polymerase (grey nucleotides), and the resulting blunt ends ligated. The mixture is subjected to high-throughput DNA sequencing. ( B ) Overhang types generated by a control restriction enzyme (HindIII) and three of the ZFN pairs used in this work. For clarity, only one DNA strand is shown. Enzyme binding sites are shown in grey; only the flanking three nucleotides are shown for ZFN binding sites. Primary cleavage sites, black triangles; secondary and tertiary cleavage sites, dark and light grey triangles, respectively; deletions, Δ. Microhomology within the target site can prevent unambiguous deduction of the overhang type. In such situations the possible overhangs are shown as joined triangles. Either of the two indicated thymidine residues may have been deleted after HindIII digestion.
    Figure Legend Snippet: Analysis of the overhang types created by ZFNs. ( A ) Scheme to determine ZFN overhangs. A supercoiled plasmid with a ZFN cleavage site is cut by a titration of in vitro transcribed and translated ZFNs. ZFN-linearized plasmids are purified by gel electrophoresis, 5′ overhangs filled in with Klenow polymerase (grey nucleotides), and the resulting blunt ends ligated. The mixture is subjected to high-throughput DNA sequencing. ( B ) Overhang types generated by a control restriction enzyme (HindIII) and three of the ZFN pairs used in this work. For clarity, only one DNA strand is shown. Enzyme binding sites are shown in grey; only the flanking three nucleotides are shown for ZFN binding sites. Primary cleavage sites, black triangles; secondary and tertiary cleavage sites, dark and light grey triangles, respectively; deletions, Δ. Microhomology within the target site can prevent unambiguous deduction of the overhang type. In such situations the possible overhangs are shown as joined triangles. Either of the two indicated thymidine residues may have been deleted after HindIII digestion.

    Techniques Used: Plasmid Preparation, Titration, In Vitro, Purification, Nucleic Acid Electrophoresis, High Throughput Screening Assay, DNA Sequencing, Generated, Binding Assay

    Related Articles

    Incubation:

    Article Title: Rpn (YhgA-Like) Proteins of Escherichia coli K-12 and Their Contribution to RecA-Independent Horizontal Transfer
    Article Snippet: .. DNA smears (1 μg) were incubated with dTTP, dATP, fluorescein–12-dCTP (catalog number NEL434001EA; PerkinElmer), fluorescein–12-dGTP (catalog number NEL496001EA; PerkinElmer), and the E. coli polymerase I Klenow fragment (catalog number M0210S; NEB) at 37°C for 30 min. Purified DNA was run on a 6% TBE-polyacrylamide gel; labeling was visualized using a Typhoon 9400 laser scanner (excitation wavelength, 488 nm; emission wavelength, 520 nm; GE Healthcare Life Sciences). .. Total DNA was visualized by UV after EtBr staining.

    Article Title: Antigenic variation by switching inter-chromosomal interactions with an RNA splicing locus in trypanosomes
    Article Snippet: .. For end-repair and biotin removal from un-ligated ends, 70 μl of end-repair mix was added (1× Ligation buffer (NEB), 357 μM dNTPs, 25U T4 PNK (NEB, M0201), 7.5U T4 DNA polymerase I (NEB, M0203), 2.5U DNA polymerase I, large (Klenow) fragment (NEB, M0210)) and incubated for 30 min at 20 °C and 20 min at 75 °C. .. To inactivate the enzymes, EDTA was added to a final concentration of 10 mM.

    other:

    Article Title: An Efficient Strategy for Broad-Range Detection of Low Abundance Bacteria without DNA Decontamination of PCR Reagents
    Article Snippet: Materials The exo I and Klenow DNA polymerase were purchased from New England Biolab (Ipswich, MA).

    Article Title: Construction of a circular single-stranded DNA template containing a defined lesion
    Article Snippet: E.coli DNA polymerase I (Klenow Fragment) [pol I (Kf)], E.coli Exonuclease III, E.coli Exonuclease I, E.coli Uracil DNA glycosylase, E.coli RecA, T7 DNA polymerase, T4 Polynucleotide kinase, and M13KO7 helper phage were all purchased from New England Biolabs (Ipswich, MA).

    Purification:

    Article Title: Rpn (YhgA-Like) Proteins of Escherichia coli K-12 and Their Contribution to RecA-Independent Horizontal Transfer
    Article Snippet: .. DNA smears (1 μg) were incubated with dTTP, dATP, fluorescein–12-dCTP (catalog number NEL434001EA; PerkinElmer), fluorescein–12-dGTP (catalog number NEL496001EA; PerkinElmer), and the E. coli polymerase I Klenow fragment (catalog number M0210S; NEB) at 37°C for 30 min. Purified DNA was run on a 6% TBE-polyacrylamide gel; labeling was visualized using a Typhoon 9400 laser scanner (excitation wavelength, 488 nm; emission wavelength, 520 nm; GE Healthcare Life Sciences). .. Total DNA was visualized by UV after EtBr staining.

    Labeling:

    Article Title: Rpn (YhgA-Like) Proteins of Escherichia coli K-12 and Their Contribution to RecA-Independent Horizontal Transfer
    Article Snippet: .. DNA smears (1 μg) were incubated with dTTP, dATP, fluorescein–12-dCTP (catalog number NEL434001EA; PerkinElmer), fluorescein–12-dGTP (catalog number NEL496001EA; PerkinElmer), and the E. coli polymerase I Klenow fragment (catalog number M0210S; NEB) at 37°C for 30 min. Purified DNA was run on a 6% TBE-polyacrylamide gel; labeling was visualized using a Typhoon 9400 laser scanner (excitation wavelength, 488 nm; emission wavelength, 520 nm; GE Healthcare Life Sciences). .. Total DNA was visualized by UV after EtBr staining.

    Ligation:

    Article Title: Antigenic variation by switching inter-chromosomal interactions with an RNA splicing locus in trypanosomes
    Article Snippet: .. For end-repair and biotin removal from un-ligated ends, 70 μl of end-repair mix was added (1× Ligation buffer (NEB), 357 μM dNTPs, 25U T4 PNK (NEB, M0201), 7.5U T4 DNA polymerase I (NEB, M0203), 2.5U DNA polymerase I, large (Klenow) fragment (NEB, M0210)) and incubated for 30 min at 20 °C and 20 min at 75 °C. .. To inactivate the enzymes, EDTA was added to a final concentration of 10 mM.

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  • 90
    New England Biolabs klenow large fragment
    Primer extension using AHP dUTP (1.5 h). ( A ) Template T2 and primer P3. ( B ) Twenty percent denaturing PAGE analysis of reactions using Gotaq (Go, 72°C), <t>Klenow</t> (Kl, 37°C), KOD (KO, 72°C) and <t>Therminator™</t> II (Th, 72°C) polymerases. Lane P: primer P3. ( C ) Reactions using Gotaq polymerase at 60°C. ( D ) Mass spectrum of AHP-modified fully extended product using Klenow (calculated mass: 10621).
    Klenow Large Fragment, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 90/100, based on 66 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/klenow large fragment/product/New England Biolabs
    Average 90 stars, based on 66 article reviews
    Price from $9.99 to $1999.99
    klenow large fragment - by Bioz Stars, 2020-07
    90/100 stars
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    99
    New England Biolabs dna polymerase i
    PAGE analysis of primer extension experiments with single OXP-modified and PDE primers. Primer extension with Klenow fragment of <t>DNA</t> <t>polymerase</t> I of nonheated ( A ) and preheated ( B ) single OXP-modified reverse primer, respectively along template 2. The extension reactions were incubated at 25°C for the indicated times after which the reaction mixtures were quenched and analyzed. ( C ) Primer extension with Taq DNA polymerase of PDE and OXP forward primers (nonheated control and preheated sample) along template oligonucleotide 1. Extension reactions were incubated at 25°C for 15 min, after which the aliquots from reaction mixtures were quenched and analyzed.
    Dna Polymerase I, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 85 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dna polymerase i/product/New England Biolabs
    Average 99 stars, based on 85 article reviews
    Price from $9.99 to $1999.99
    dna polymerase i - by Bioz Stars, 2020-07
    99/100 stars
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    Image Search Results


    Primer extension using AHP dUTP (1.5 h). ( A ) Template T2 and primer P3. ( B ) Twenty percent denaturing PAGE analysis of reactions using Gotaq (Go, 72°C), Klenow (Kl, 37°C), KOD (KO, 72°C) and Therminator™ II (Th, 72°C) polymerases. Lane P: primer P3. ( C ) Reactions using Gotaq polymerase at 60°C. ( D ) Mass spectrum of AHP-modified fully extended product using Klenow (calculated mass: 10621).

    Journal: Nucleic Acids Research

    Article Title: Efficient enzymatic synthesis and dual-colour fluorescent labelling of DNA probes using long chain azido-dUTP and BCN dyes

    doi: 10.1093/nar/gkw028

    Figure Lengend Snippet: Primer extension using AHP dUTP (1.5 h). ( A ) Template T2 and primer P3. ( B ) Twenty percent denaturing PAGE analysis of reactions using Gotaq (Go, 72°C), Klenow (Kl, 37°C), KOD (KO, 72°C) and Therminator™ II (Th, 72°C) polymerases. Lane P: primer P3. ( C ) Reactions using Gotaq polymerase at 60°C. ( D ) Mass spectrum of AHP-modified fully extended product using Klenow (calculated mass: 10621).

    Article Snippet: Klenow large fragment, Therminator™ II, M-MuLV (RNase H− ) reverse transcriptase, AMV reverse transcriptase, RNase inhibitor and λ-exonuclease were purchased from New England Biolabs.

    Techniques: Polyacrylamide Gel Electrophoresis, Modification

    UL30 inhibits the minicircle replication in the absence of UL42 Reactions contained helicase, polymerase(s), DNA MC70-2 (A) and were quenched after 30 minutes. (B) Lanes 1–6 contained 100 nM Klenow Fragment and increasing concentrations of UL30 (0, 10, 50, 100, 150 or 200 nM). Lanes 7–12 contained 100 nM UL30 and increasing concentrations of Klenow Fragment (0, 10, 50, 100, 150 or 200 nM). DNA products were separated using 1.5% alkaline agarose gel electrophoresis. (C) Amount of dNTPs incorporated was measured using ImageQuant.

    Journal: Biochemistry

    Article Title: Protein Displacement by Herpes Helicase-Primase and the Key Role of UL42 During Helicase-Coupled DNA Synthesis by the Herpes Polymerase

    doi: 10.1021/acs.biochem.6b01128

    Figure Lengend Snippet: UL30 inhibits the minicircle replication in the absence of UL42 Reactions contained helicase, polymerase(s), DNA MC70-2 (A) and were quenched after 30 minutes. (B) Lanes 1–6 contained 100 nM Klenow Fragment and increasing concentrations of UL30 (0, 10, 50, 100, 150 or 200 nM). Lanes 7–12 contained 100 nM UL30 and increasing concentrations of Klenow Fragment (0, 10, 50, 100, 150 or 200 nM). DNA products were separated using 1.5% alkaline agarose gel electrophoresis. (C) Amount of dNTPs incorporated was measured using ImageQuant.

    Article Snippet: Both polymerases could replace UL30-UL42, with Klenow Fragment generating products ~1 kB long.

    Techniques: Agarose Gel Electrophoresis

    Non-cognate polymerases can replace UL30-UL42 during minicircle replication Either Klenow Fragment or T4 DNA Polymerase were titrated into assays containing DNA MC70 (A) and 100 nM UL5-UL8-UL52. (B) DNA products were separated with 1.5% alkaline agarose gel electrophoresis.

    Journal: Biochemistry

    Article Title: Protein Displacement by Herpes Helicase-Primase and the Key Role of UL42 During Helicase-Coupled DNA Synthesis by the Herpes Polymerase

    doi: 10.1021/acs.biochem.6b01128

    Figure Lengend Snippet: Non-cognate polymerases can replace UL30-UL42 during minicircle replication Either Klenow Fragment or T4 DNA Polymerase were titrated into assays containing DNA MC70 (A) and 100 nM UL5-UL8-UL52. (B) DNA products were separated with 1.5% alkaline agarose gel electrophoresis.

    Article Snippet: Both polymerases could replace UL30-UL42, with Klenow Fragment generating products ~1 kB long.

    Techniques: Agarose Gel Electrophoresis

    PAGE analysis of primer extension experiments with single OXP-modified and PDE primers. Primer extension with Klenow fragment of DNA polymerase I of nonheated ( A ) and preheated ( B ) single OXP-modified reverse primer, respectively along template 2. The extension reactions were incubated at 25°C for the indicated times after which the reaction mixtures were quenched and analyzed. ( C ) Primer extension with Taq DNA polymerase of PDE and OXP forward primers (nonheated control and preheated sample) along template oligonucleotide 1. Extension reactions were incubated at 25°C for 15 min, after which the aliquots from reaction mixtures were quenched and analyzed.

    Journal: Nucleic Acids Research

    Article Title: Hot Start PCR with heat-activatable primers: a novel approach for improved PCR performance

    doi: 10.1093/nar/gkn575

    Figure Lengend Snippet: PAGE analysis of primer extension experiments with single OXP-modified and PDE primers. Primer extension with Klenow fragment of DNA polymerase I of nonheated ( A ) and preheated ( B ) single OXP-modified reverse primer, respectively along template 2. The extension reactions were incubated at 25°C for the indicated times after which the reaction mixtures were quenched and analyzed. ( C ) Primer extension with Taq DNA polymerase of PDE and OXP forward primers (nonheated control and preheated sample) along template oligonucleotide 1. Extension reactions were incubated at 25°C for 15 min, after which the aliquots from reaction mixtures were quenched and analyzed.

    Article Snippet: Primer extension with Klenow fragment of DNA polymerase Primer extension experiments using large fragment (Klenow) of DNA polymerase I (New England Biolabs) were performed at 25°C using the HIV-1 tat reverse primer (5′-AATACTATGGTCCACACAACTATTGCT-3′) that was unmodified or contained a single OXP modification.

    Techniques: Polyacrylamide Gel Electrophoresis, Modification, Incubation