klenow dna polymerase  (New England Biolabs)


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    Name:
    DNA Polymerase I Klenow Fragment
    Description:
    DNA Polymerase I Klenow Fragment 1 000 units
    Catalog Number:
    m0210l
    Price:
    248
    Category:
    DNA Polymerases
    Size:
    1 000 units
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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 1 article reviews
    Price from $9.99 to $1999.99
    klenow dna polymerase - by Bioz Stars, 2021-03
    99/100 stars

    Images

    1) 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

    Related Articles

    other:

    Article Title: Hot Start PCR with heat-activatable primers: a novel approach for improved PCR performance
    Article Snippet: Extension of PTE primers by DNA polymerases The ability of Klenow fragment of DNA polymerase I and Taq DNA polymerase to perform template-dependent extension and elongation of the HIV-1 single OXP-modified primers was investigated.

    Article Title: Protein Displacement by Herpes Helicase-Primase and the Key Role of UL42 During Helicase-Coupled DNA Synthesis by the Herpes Polymerase
    Article Snippet: Both polymerases could replace UL30-UL42, with Klenow Fragment generating products ~1 kB long.

    Article Title: Protein Displacement by Herpes Helicase-Primase and the Key Role of UL42 During Helicase-Coupled DNA Synthesis by the Herpes Polymerase
    Article Snippet: The absence of any products longer than ~100 nucleotides in assays containing UL30 is not due to either the lower processivity of UL30 relative to UL30-UL42 since UL30 and Klenow Fragment have similar processivity ( ) and Klenow Fragment generates long products.

    Article Title: Cleavage of deoxyoxanosine-containing oligodeoxyribonucleotides by bacterial endonuclease V
    Article Snippet: Klenow Fragment (3′–5′ exo-) was purchased from New England Biolabs (Beverly, MA).

    Article Title: Protein Displacement by Herpes Helicase-Primase and the Key Role of UL42 During Helicase-Coupled DNA Synthesis by the Herpes Polymerase
    Article Snippet: Even without ICP8, we found that non-cognate polymerases such as Klenow Fragment and T4 DNA polymerase allow the herpes helicase to more efficiently unwind DNA and allow the synthesis of long products.

    Modification:

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

    DNA Synthesis:

    Article Title: 5?CAG and 5?CTG Repeats Create Differential Impediment to the Progression of a Minimal Reconstituted T4 Replisome Depending on the Concentration of dNTPs
    Article Snippet: Klenow fragment has weak strand displacement DNA synthesis activity that allowed it to synthesize through the duplex part of the minifork ( , lanes 2 and 5). .. However, contrary to the gp43-gp41 replication couple, DNA synthesis performed by Klenow fragment was not stimulated by gp41 ( , compare lanes 2, 3, 5, and 6). .. Under both conditions (with or without gp41), DNA synthesis was highly distributive as indicated by the numerous pause sites that were clearly visible along the template strand.

    Plasmid Preparation:

    Article Title: Establishment of conditional vectors for hairpin siRNA knockdowns
    Article Snippet: The human U6 promoter was amplified by PCR from the genomic DNA of T24 cells with the primers U6proTop (5′-AGGCAAAACGCACCACGTGACGGAG-3′) and U6proBottomSphI (5′-GCATGCGGTGTTTCGTCCTTTCCACAAGAT-3′), then ligated into a TA cloning vector, pGEM-T Easy (Promega, Madison, WI) (Fig. B). .. The plasmid was then digested with SphI (New England Biolabs, Beverly, MA) and blunted with Klenow fragment (New England Biolabs) (Fig. C). .. Oligo1 [a mixture of two DNA oligomers, DNMTO1F (5′-GGAATGGCAGATGCCAACAGGACGT-3′) and DNMTO1R (5′-CCTGTTGGCATCTGCCATTCC-3′)] was ligated following restriction digestion with AatII (New England Biolabs) (Fig. D).

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    New England Biolabs escherichia coli dna polymerase i
    DSBs ( a ) and SSBs ( b – d ) generated in presence of Top2 ( a ); ETO ( b ); Top1 ( c ); and <t>DNA-damaging</t> agents that modify the DNA termini ( d ). Red arrow represents successful nick translation. Stop sign represents unsuccessful nick translation. Nick translation by DNA <t>polymerase</t> I necessitates a 3'-OH, which is not reconstituted in case of Top1 cleavage or when the DNA termini is damaged (shown by asterisk). In these cases the principal enzymes involved in processing and repair of the ends are listed below the black arrow. TDP1, tyrosyl-DNA phosphodiesterase 1, PNKP, polynucleotide kinase 3'-phosphatase, APE1, AP endonuclease I [ 17 ]
    Escherichia Coli Dna Polymerase I, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/escherichia coli dna polymerase i/product/New England Biolabs
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
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    99
    New England Biolabs klenow fragment
    Schematic overview of random deletion used in this study . A) Linearization of the plasmid by restriction digestion at Sca I site. B) Random deletion with Exo III. C) Blunt-ended with mung bean nuclease and <t>Klenow</t> fragment. D) Single digested with Bgl II. E) Random ligation with T4 <t>DNA</t> ligase.
    Klenow Fragment, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/klenow fragment/product/New England Biolabs
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    klenow fragment - by Bioz Stars, 2021-03
    99/100 stars
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    Image Search Results


    DSBs ( a ) and SSBs ( b – d ) generated in presence of Top2 ( a ); ETO ( b ); Top1 ( c ); and DNA-damaging agents that modify the DNA termini ( d ). Red arrow represents successful nick translation. Stop sign represents unsuccessful nick translation. Nick translation by DNA polymerase I necessitates a 3'-OH, which is not reconstituted in case of Top1 cleavage or when the DNA termini is damaged (shown by asterisk). In these cases the principal enzymes involved in processing and repair of the ends are listed below the black arrow. TDP1, tyrosyl-DNA phosphodiesterase 1, PNKP, polynucleotide kinase 3'-phosphatase, APE1, AP endonuclease I [ 17 ]

    Journal: International Journal of Molecular Sciences

    Article Title: DNA Break Mapping Reveals Topoisomerase II Activity Genome-Wide

    doi: 10.3390/ijms150713111

    Figure Lengend Snippet: DSBs ( a ) and SSBs ( b – d ) generated in presence of Top2 ( a ); ETO ( b ); Top1 ( c ); and DNA-damaging agents that modify the DNA termini ( d ). Red arrow represents successful nick translation. Stop sign represents unsuccessful nick translation. Nick translation by DNA polymerase I necessitates a 3'-OH, which is not reconstituted in case of Top1 cleavage or when the DNA termini is damaged (shown by asterisk). In these cases the principal enzymes involved in processing and repair of the ends are listed below the black arrow. TDP1, tyrosyl-DNA phosphodiesterase 1, PNKP, polynucleotide kinase 3'-phosphatase, APE1, AP endonuclease I [ 17 ]

    Article Snippet: 500 μg of DNA was incubated for 40 s at 16 °C with a mixture of 200 μM of dATP, dGTP, dCTP and 20 μM of digoxigenin-11-dUTP (Roche), 117 μM of ddATP, ddGTP, ddCTP (Roche) and 1000 units of Escherichia coli DNA Polymerase I (New England Biolabs, Ipswich, MA, USA).

    Techniques: Generated, Nick Translation

    Schematic overview of random deletion used in this study . A) Linearization of the plasmid by restriction digestion at Sca I site. B) Random deletion with Exo III. C) Blunt-ended with mung bean nuclease and Klenow fragment. D) Single digested with Bgl II. E) Random ligation with T4 DNA ligase.

    Journal: Microbial Cell Factories

    Article Title: An evolved xylose transporter from Zymomonas mobilis enhances sugar transport in Escherichia coli

    doi: 10.1186/1475-2859-8-66

    Figure Lengend Snippet: Schematic overview of random deletion used in this study . A) Linearization of the plasmid by restriction digestion at Sca I site. B) Random deletion with Exo III. C) Blunt-ended with mung bean nuclease and Klenow fragment. D) Single digested with Bgl II. E) Random ligation with T4 DNA ligase.

    Article Snippet: To facilitate the re-ligation (see below), the purified DNA was blunt-ended with 3.2 U Klenow fragment in 10× NEBuffer 2 (New England Biolabs) and dNTPs (final concentration 33 μM each nucleotide) for 15 min at 25°C.

    Techniques: Plasmid Preparation, Ligation

    Overview of the experimental steps required to create and analyse a chromatin accessibility library. ( A ) Step 1: fungal mycelia pre-grown under specific conditions or isolated DNA ( in vitro controls) are processed as described in Materials and methods section and digested with MNase or restriction enzymes of choice. Step 2: digested DNA is blunt-ended and phosphorylated by subsequent treatment of the chromatin with Klenow fragment polymerase, T4 polynucleotide kinase. This step produces blunt-ended DNA fragments for ligation with adaptors. Step 3: DNA fragments are ligated with double-stranded adaptors A and B, originating from oligonucleotides Adaptor-A short and Adaptor-A long or Adaptor-B short and Adaptor-B long , where adaptor oligonucleotide B long is biotinylated for later retention on the streptavidin beads. In this step, fragments containing all adaptor combinations (A-A, A-B and B-B) are generated. Step 4: the ligation step leaves nicks at the 3′-terminus that are repaired by Bst polymerase treatment. Step 5: all fragments containing biotinylated adaptor B are captured on streptavidin-coated magnetic beads. At this step, adaptor A-A fragments are lost. Step 6: after a washing step, the retained fragments (adaptors A-B and B-B fragments) are denatured at 95°C. The denaturation step results in the release of single strands which exclusively carry A-B adaptor fragments. Step 7: the single-stranded A-B adaptor fragment library is amplified by a nested PCR approach to give the final A-B fragment library. The input and output fragment libraries are quality controlled by amplification with single A and B, as well as mixed A-B primers. Only the A-B primer mix should result in the amplification of fragments in the range of 200–1000 bp (see Panel B). Step 8: the resulting A-B adaptor fragment library is diluted and aliquots are used for analytical PCR amplifications for fragment size analysis of specific loci of interest. In the final analytical PCR step, either gene-specific or adaptor-specific primers can be labelled for subsequent capillary sequencer analysis. The chromatograms are finally analysed by image analysis software. ( B ) Example of quality control of A-B adaptor fragment libraries. Two input chromatin fragment libraries without adaptor ligation (lanes 1 and 2) are compared to two output libraries with adaptor ligation as described in Materials and methods section (lanes 3 and 4). Libraries originating from nitrate-grown cells (lanes 1 and 3) as well as from ammonium-grown cells (lanes 2 and 4) are shown as an example. M, DNA size marker.

    Journal: Nucleic Acids Research

    Article Title: A library-based method to rapidly analyse chromatin accessibility at multiple genomic regions

    doi: 10.1093/nar/gkp037

    Figure Lengend Snippet: Overview of the experimental steps required to create and analyse a chromatin accessibility library. ( A ) Step 1: fungal mycelia pre-grown under specific conditions or isolated DNA ( in vitro controls) are processed as described in Materials and methods section and digested with MNase or restriction enzymes of choice. Step 2: digested DNA is blunt-ended and phosphorylated by subsequent treatment of the chromatin with Klenow fragment polymerase, T4 polynucleotide kinase. This step produces blunt-ended DNA fragments for ligation with adaptors. Step 3: DNA fragments are ligated with double-stranded adaptors A and B, originating from oligonucleotides Adaptor-A short and Adaptor-A long or Adaptor-B short and Adaptor-B long , where adaptor oligonucleotide B long is biotinylated for later retention on the streptavidin beads. In this step, fragments containing all adaptor combinations (A-A, A-B and B-B) are generated. Step 4: the ligation step leaves nicks at the 3′-terminus that are repaired by Bst polymerase treatment. Step 5: all fragments containing biotinylated adaptor B are captured on streptavidin-coated magnetic beads. At this step, adaptor A-A fragments are lost. Step 6: after a washing step, the retained fragments (adaptors A-B and B-B fragments) are denatured at 95°C. The denaturation step results in the release of single strands which exclusively carry A-B adaptor fragments. Step 7: the single-stranded A-B adaptor fragment library is amplified by a nested PCR approach to give the final A-B fragment library. The input and output fragment libraries are quality controlled by amplification with single A and B, as well as mixed A-B primers. Only the A-B primer mix should result in the amplification of fragments in the range of 200–1000 bp (see Panel B). Step 8: the resulting A-B adaptor fragment library is diluted and aliquots are used for analytical PCR amplifications for fragment size analysis of specific loci of interest. In the final analytical PCR step, either gene-specific or adaptor-specific primers can be labelled for subsequent capillary sequencer analysis. The chromatograms are finally analysed by image analysis software. ( B ) Example of quality control of A-B adaptor fragment libraries. Two input chromatin fragment libraries without adaptor ligation (lanes 1 and 2) are compared to two output libraries with adaptor ligation as described in Materials and methods section (lanes 3 and 4). Libraries originating from nitrate-grown cells (lanes 1 and 3) as well as from ammonium-grown cells (lanes 2 and 4) are shown as an example. M, DNA size marker.

    Article Snippet: To allow for ligation with the blunt-end of the double-stranded adaptors, 15 µl of purified MNase digested DNA fragments were blunt-ended in a final volume of 20 µl by Klenow fragment of Escherichia coli DNA polymerase (New England Biolabs, CA, USA) and phosphorylated by T4 polynucleotide kinase (New England Biolabs) in a final volume of 30 µl.

    Techniques: Isolation, In Vitro, Ligation, Generated, Magnetic Beads, Amplification, Nested PCR, Polymerase Chain Reaction, Software, Marker