therminator dna polymerase  (New England Biolabs)


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    New England Biolabs therminator dna polymerase
    Therminator Dna Polymerase, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    therminator dna polymerase  (New England Biolabs)


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    New England Biolabs therminator dna polymerase
    Therminator Dna Polymerase, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    therminator dna polymerase  (New England Biolabs)


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    New England Biolabs therminator dna polymerase
    TNA polymerization fidelity by position and sequence
    Therminator Dna Polymerase, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "High fidelity TNA synthesis by Therminator polymerase"

    Article Title: High fidelity TNA synthesis by Therminator polymerase

    Journal: Nucleic Acids Research

    doi: 10.1093/nar/gki840

    TNA polymerization fidelity by position and sequence
    Figure Legend Snippet: TNA polymerization fidelity by position and sequence

    Techniques Used:

    therminator dna polymerase  (New England Biolabs)


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    New England Biolabs therminator dna polymerase
    Polyacrylamide gel time course of LNA-modified primer extension by 0.2 units of exo– A488L Vent <t>DNA</t> polymerase with aminoallyl-dUTP, followed by exo III digestion. (A) PAI-L1. (B) PAI-L1,2. Lanes: M, 25 bp ladder; 1, 0 min; 2, 15 min; 3, 30 min; 4, 60 min; 5, 120 min; 6, 240 min; 7, 960 min; and C, undigested oligonucleotide control.
    Therminator Dna Polymerase, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Strong positional preference in the interaction of LNA oligonucleotides with DNA polymerase and proofreading exonuclease activities: implications for genotyping assays"

    Article Title: Strong positional preference in the interaction of LNA oligonucleotides with DNA polymerase and proofreading exonuclease activities: implications for genotyping assays

    Journal:

    doi: 10.1093/nar/gnh036

    Polyacrylamide gel time course of LNA-modified primer extension by 0.2 units of exo– A488L Vent DNA polymerase with aminoallyl-dUTP, followed by exo III digestion. (A) PAI-L1. (B) PAI-L1,2. Lanes: M, 25 bp ladder; 1, 0 min; 2, 15 min; 3, 30 min; 4, 60 min; 5, 120 min; 6, 240 min; 7, 960 min; and C, undigested oligonucleotide control.
    Figure Legend Snippet: Polyacrylamide gel time course of LNA-modified primer extension by 0.2 units of exo– A488L Vent DNA polymerase with aminoallyl-dUTP, followed by exo III digestion. (A) PAI-L1. (B) PAI-L1,2. Lanes: M, 25 bp ladder; 1, 0 min; 2, 15 min; 3, 30 min; 4, 60 min; 5, 120 min; 6, 240 min; 7, 960 min; and C, undigested oligonucleotide control.

    Techniques Used: Modification

    Polymerase-mediated extension of LNA-modified primers with dTTP by (A) exo– Klenow DNA polymerase and (B) exo– A488L Vent DNA polymerase. Oligonucleotides are PAI-L1 (squares) and PAI-L1,2 (inverted triangles).
    Figure Legend Snippet: Polymerase-mediated extension of LNA-modified primers with dTTP by (A) exo– Klenow DNA polymerase and (B) exo– A488L Vent DNA polymerase. Oligonucleotides are PAI-L1 (squares) and PAI-L1,2 (inverted triangles).

    Techniques Used: Modification

    Demonstration of PRASE PCR. (A) Forward allele-specific primers and template sequences. (B) PCR amplicons generated with exo– Vent DNA polymerase. (C) PCR amplicons generated with exo+ Vent DNA polymerase. Lanes: M, 100 bp marker ladder and PCR amplicons generated with forward primers. Standard primers: 1, PAP-1g; 2, PAP-1a; 3, PAP-2a and 4, PAP-2g; L-1 LNA-modified primers: 5, PAP-1gL1; 6, PAP-1aL1; 7, PAP-2aL1; 8, PAP-2gL1; and L-2 LNA-modified primers: 9, PAP-1gL2; 10, PAP-1aL2; 11, PAP-2aL2; and 12, PAP-2gL2.
    Figure Legend Snippet: Demonstration of PRASE PCR. (A) Forward allele-specific primers and template sequences. (B) PCR amplicons generated with exo– Vent DNA polymerase. (C) PCR amplicons generated with exo+ Vent DNA polymerase. Lanes: M, 100 bp marker ladder and PCR amplicons generated with forward primers. Standard primers: 1, PAP-1g; 2, PAP-1a; 3, PAP-2a and 4, PAP-2g; L-1 LNA-modified primers: 5, PAP-1gL1; 6, PAP-1aL1; 7, PAP-2aL1; 8, PAP-2gL1; and L-2 LNA-modified primers: 9, PAP-1gL2; 10, PAP-1aL2; 11, PAP-2aL2; and 12, PAP-2gL2.

    Techniques Used: Generated, Marker, Modification

    therminator polymerase  (New England Biolabs)


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    New England Biolabs therminator polymerase
    Template-directed assembly of Z-TFOs. ( A ) Strategy for the assembly of oligonucleotides containing a triplex-forming sequence (TFS; shown in red) containing Z. (i) Primer extension of a template that contains the W–C complement to the TFO (underlined) allows the incorporation of Z by the formation of a GZ base pair at high pH (and the absence of dCTP). (ii) The modified oligonucleotide can then be isolated by selective degradation of the phosphate-labelled template (in yellow) by the action of lambda exonuclease. ( B ) Electrophoretic mobility shift assay showing the products of the extension and digestion of such reactions. The composition of each sample is shown above each lane of the gel. Final concentration of the strands and dNTPS was 5 and 100 μM, respectively. Extension reactions were performed for 2 h at 72°C with 2 units of <t>Therminator</t> Pol, whilst digestion reactions were performed for 2 h with 10 units of Lambda exonuclease at 37°C. The complexes were then separated on a 15% non-denaturing polyacrylamide gel and subjected to post-staining with GelRed.
    Therminator Polymerase, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Triplex-forming properties and enzymatic incorporation of a base-modified nucleotide capable of duplex DNA recognition at neutral pH"

    Article Title: Triplex-forming properties and enzymatic incorporation of a base-modified nucleotide capable of duplex DNA recognition at neutral pH

    Journal: Nucleic Acids Research

    doi: 10.1093/nar/gkab572

    Template-directed assembly of Z-TFOs. ( A ) Strategy for the assembly of oligonucleotides containing a triplex-forming sequence (TFS; shown in red) containing Z. (i) Primer extension of a template that contains the W–C complement to the TFO (underlined) allows the incorporation of Z by the formation of a GZ base pair at high pH (and the absence of dCTP). (ii) The modified oligonucleotide can then be isolated by selective degradation of the phosphate-labelled template (in yellow) by the action of lambda exonuclease. ( B ) Electrophoretic mobility shift assay showing the products of the extension and digestion of such reactions. The composition of each sample is shown above each lane of the gel. Final concentration of the strands and dNTPS was 5 and 100 μM, respectively. Extension reactions were performed for 2 h at 72°C with 2 units of Therminator Pol, whilst digestion reactions were performed for 2 h with 10 units of Lambda exonuclease at 37°C. The complexes were then separated on a 15% non-denaturing polyacrylamide gel and subjected to post-staining with GelRed.
    Figure Legend Snippet: Template-directed assembly of Z-TFOs. ( A ) Strategy for the assembly of oligonucleotides containing a triplex-forming sequence (TFS; shown in red) containing Z. (i) Primer extension of a template that contains the W–C complement to the TFO (underlined) allows the incorporation of Z by the formation of a GZ base pair at high pH (and the absence of dCTP). (ii) The modified oligonucleotide can then be isolated by selective degradation of the phosphate-labelled template (in yellow) by the action of lambda exonuclease. ( B ) Electrophoretic mobility shift assay showing the products of the extension and digestion of such reactions. The composition of each sample is shown above each lane of the gel. Final concentration of the strands and dNTPS was 5 and 100 μM, respectively. Extension reactions were performed for 2 h at 72°C with 2 units of Therminator Pol, whilst digestion reactions were performed for 2 h with 10 units of Lambda exonuclease at 37°C. The complexes were then separated on a 15% non-denaturing polyacrylamide gel and subjected to post-staining with GelRed.

    Techniques Used: Sequencing, Modification, Isolation, Electrophoretic Mobility Shift Assay, Concentration Assay, Staining

    dna polymerases  (New England Biolabs)


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    New England Biolabs dna polymerases
    Dna Polymerases, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    dna polymerases  (New England Biolabs)


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    New England Biolabs dna polymerases
    Dna Polymerases, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    dna polymerases  (New England Biolabs)


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    New England Biolabs dna polymerases
    Dna Polymerases, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    therminator  (New England Biolabs)


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    New England Biolabs therminator
    PAGE analysis of the incorporation of PEG modified dUTPs with the use of Vent ( exo- ), <t>Therminator,</t> Pfu and Bst 2.0 DNA polymerases. All lanes except “no dNTPs” contain dATP, dCTP, dGTP and the nucleotide indicated above the gel (from left): Lane 1 = dTTP, dATP, dCTP, dGTP; Lane 2 = PEG 8 -dUTP ( 22 ), dATP, dCTP, dGTP; Lane 3 = PEG 16 -dUTP ( 23 ), dATP, dCTP, dGTP; Lane 4 = PEG 24 -dUTP ( 24 ), dATP, dCTP, dGTP; Lane 5 = PEG 6 -dUTP, dATP, dCTP, dGTP; Lane 6 = dATP, dCTP, dGTP; Lane 7 = Control, no dNTPs present. Conditions for all primer reactions; 20 nM primer, 15 nM template, 625 nM dNTPs, 40 °C, 1 h.
    Therminator, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Synthesis of Polyanionic C5-Modified 2′-Deoxyuridine and 2′-Deoxycytidine-5′-Triphosphates and Their Properties as Substrates for DNA Polymerases"

    Article Title: Synthesis of Polyanionic C5-Modified 2′-Deoxyuridine and 2′-Deoxycytidine-5′-Triphosphates and Their Properties as Substrates for DNA Polymerases

    Journal: Molecules

    doi: 10.3390/molecules26082250

    PAGE analysis of the incorporation of PEG modified dUTPs with the use of Vent ( exo- ), Therminator, Pfu and Bst 2.0 DNA polymerases. All lanes except “no dNTPs” contain dATP, dCTP, dGTP and the nucleotide indicated above the gel (from left): Lane 1 = dTTP, dATP, dCTP, dGTP; Lane 2 = PEG 8 -dUTP ( 22 ), dATP, dCTP, dGTP; Lane 3 = PEG 16 -dUTP ( 23 ), dATP, dCTP, dGTP; Lane 4 = PEG 24 -dUTP ( 24 ), dATP, dCTP, dGTP; Lane 5 = PEG 6 -dUTP, dATP, dCTP, dGTP; Lane 6 = dATP, dCTP, dGTP; Lane 7 = Control, no dNTPs present. Conditions for all primer reactions; 20 nM primer, 15 nM template, 625 nM dNTPs, 40 °C, 1 h.
    Figure Legend Snippet: PAGE analysis of the incorporation of PEG modified dUTPs with the use of Vent ( exo- ), Therminator, Pfu and Bst 2.0 DNA polymerases. All lanes except “no dNTPs” contain dATP, dCTP, dGTP and the nucleotide indicated above the gel (from left): Lane 1 = dTTP, dATP, dCTP, dGTP; Lane 2 = PEG 8 -dUTP ( 22 ), dATP, dCTP, dGTP; Lane 3 = PEG 16 -dUTP ( 23 ), dATP, dCTP, dGTP; Lane 4 = PEG 24 -dUTP ( 24 ), dATP, dCTP, dGTP; Lane 5 = PEG 6 -dUTP, dATP, dCTP, dGTP; Lane 6 = dATP, dCTP, dGTP; Lane 7 = Control, no dNTPs present. Conditions for all primer reactions; 20 nM primer, 15 nM template, 625 nM dNTPs, 40 °C, 1 h.

    Techniques Used: Modification

    PAGE analysis of the incorporation of PEG modified dCTPs with the use of Omni-Klentaq, Therminator, Klenow ( exo- ) and Vent ( exo- ) DNA polymerase. All lanes except “no dNTPs” contain dATP, dGTP, dTTP and the nucleotide indicated above the gel (from left): Lane 1 = dCTP, dATP, dGTP, dTTP; Lane 2 = PEG 8 -dCTP ( 25 ), dATP, dGTP, dTTP; Lane 3 = PEG 4 -dCTP ( 26 ), dATP, dGTP, dTTP; Lane 4 = dATP, dGTP, dTTP; Lane 5 = Control, no dNTPs present. Conditions for all primer reactions; 20 nM primer, 15 nM template, 625 nM dNTPs, 40 °C, 1 h.
    Figure Legend Snippet: PAGE analysis of the incorporation of PEG modified dCTPs with the use of Omni-Klentaq, Therminator, Klenow ( exo- ) and Vent ( exo- ) DNA polymerase. All lanes except “no dNTPs” contain dATP, dGTP, dTTP and the nucleotide indicated above the gel (from left): Lane 1 = dCTP, dATP, dGTP, dTTP; Lane 2 = PEG 8 -dCTP ( 25 ), dATP, dGTP, dTTP; Lane 3 = PEG 4 -dCTP ( 26 ), dATP, dGTP, dTTP; Lane 4 = dATP, dGTP, dTTP; Lane 5 = Control, no dNTPs present. Conditions for all primer reactions; 20 nM primer, 15 nM template, 625 nM dNTPs, 40 °C, 1 h.

    Techniques Used: Modification

    therminator dna polymerase  (New England Biolabs)


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    New England Biolabs therminator dna polymerase
    Therminator Dna Polymerase, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    therminator ix  (New England Biolabs)


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    New England Biolabs therminator ix
    Incorporation of 2’‐F,Me‐UTP as a terminator by two low‐fidelity polymerases but not by a high‐fidelity polymerase. The sequence of the primer and template used for these extension reactions is shown at the top of the figure. Polymerase extension reactions were performed by incubating the primer and template with 2’‐F,Me‐UTP and the appropriate reaction buffer for the specific enzyme, followed by detection of the reaction products by MALDI‐TOF MS. The MS spectra of the extension products generated by <t>Therminator</t> II (T2) in (A) and Therminator IX (T9) in (B) indicate single‐base incorporation and termination, whereas the MS spectrum for Thermo Sequenase (TS) in (C) indicates no incorporation, showing only a primer peak. The accuracy for m/z determination is ± 10 Da
    Therminator Ix, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Nucleotide analogues as inhibitors of SARS‐CoV Polymerase"

    Article Title: Nucleotide analogues as inhibitors of SARS‐CoV Polymerase

    Journal: Pharmacology Research & Perspectives

    doi: 10.1002/prp2.674

    Incorporation of 2’‐F,Me‐UTP as a terminator by two low‐fidelity polymerases but not by a high‐fidelity polymerase. The sequence of the primer and template used for these extension reactions is shown at the top of the figure. Polymerase extension reactions were performed by incubating the primer and template with 2’‐F,Me‐UTP and the appropriate reaction buffer for the specific enzyme, followed by detection of the reaction products by MALDI‐TOF MS. The MS spectra of the extension products generated by Therminator II (T2) in (A) and Therminator IX (T9) in (B) indicate single‐base incorporation and termination, whereas the MS spectrum for Thermo Sequenase (TS) in (C) indicates no incorporation, showing only a primer peak. The accuracy for m/z determination is ± 10 Da
    Figure Legend Snippet: Incorporation of 2’‐F,Me‐UTP as a terminator by two low‐fidelity polymerases but not by a high‐fidelity polymerase. The sequence of the primer and template used for these extension reactions is shown at the top of the figure. Polymerase extension reactions were performed by incubating the primer and template with 2’‐F,Me‐UTP and the appropriate reaction buffer for the specific enzyme, followed by detection of the reaction products by MALDI‐TOF MS. The MS spectra of the extension products generated by Therminator II (T2) in (A) and Therminator IX (T9) in (B) indicate single‐base incorporation and termination, whereas the MS spectrum for Thermo Sequenase (TS) in (C) indicates no incorporation, showing only a primer peak. The accuracy for m/z determination is ± 10 Da

    Techniques Used: Sequencing, Generated

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    New England Biolabs therminator dna polymerase
    Therminator Dna Polymerase, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    New England Biolabs therminator polymerase
    Template-directed assembly of Z-TFOs. ( A ) Strategy for the assembly of oligonucleotides containing a triplex-forming sequence (TFS; shown in red) containing Z. (i) Primer extension of a template that contains the W–C complement to the TFO (underlined) allows the incorporation of Z by the formation of a GZ base pair at high pH (and the absence of dCTP). (ii) The modified oligonucleotide can then be isolated by selective degradation of the phosphate-labelled template (in yellow) by the action of lambda exonuclease. ( B ) Electrophoretic mobility shift assay showing the products of the extension and digestion of such reactions. The composition of each sample is shown above each lane of the gel. Final concentration of the strands and dNTPS was 5 and 100 μM, respectively. Extension reactions were performed for 2 h at 72°C with 2 units of <t>Therminator</t> Pol, whilst digestion reactions were performed for 2 h with 10 units of Lambda exonuclease at 37°C. The complexes were then separated on a 15% non-denaturing polyacrylamide gel and subjected to post-staining with GelRed.
    Therminator Polymerase, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    New England Biolabs dna polymerases
    Template-directed assembly of Z-TFOs. ( A ) Strategy for the assembly of oligonucleotides containing a triplex-forming sequence (TFS; shown in red) containing Z. (i) Primer extension of a template that contains the W–C complement to the TFO (underlined) allows the incorporation of Z by the formation of a GZ base pair at high pH (and the absence of dCTP). (ii) The modified oligonucleotide can then be isolated by selective degradation of the phosphate-labelled template (in yellow) by the action of lambda exonuclease. ( B ) Electrophoretic mobility shift assay showing the products of the extension and digestion of such reactions. The composition of each sample is shown above each lane of the gel. Final concentration of the strands and dNTPS was 5 and 100 μM, respectively. Extension reactions were performed for 2 h at 72°C with 2 units of <t>Therminator</t> Pol, whilst digestion reactions were performed for 2 h with 10 units of Lambda exonuclease at 37°C. The complexes were then separated on a 15% non-denaturing polyacrylamide gel and subjected to post-staining with GelRed.
    Dna Polymerases, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    New England Biolabs therminator
    PAGE analysis of the incorporation of PEG modified dUTPs with the use of Vent ( exo- ), <t>Therminator,</t> Pfu and Bst 2.0 DNA polymerases. All lanes except “no dNTPs” contain dATP, dCTP, dGTP and the nucleotide indicated above the gel (from left): Lane 1 = dTTP, dATP, dCTP, dGTP; Lane 2 = PEG 8 -dUTP ( 22 ), dATP, dCTP, dGTP; Lane 3 = PEG 16 -dUTP ( 23 ), dATP, dCTP, dGTP; Lane 4 = PEG 24 -dUTP ( 24 ), dATP, dCTP, dGTP; Lane 5 = PEG 6 -dUTP, dATP, dCTP, dGTP; Lane 6 = dATP, dCTP, dGTP; Lane 7 = Control, no dNTPs present. Conditions for all primer reactions; 20 nM primer, 15 nM template, 625 nM dNTPs, 40 °C, 1 h.
    Therminator, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    New England Biolabs therminator ix
    Incorporation of 2’‐F,Me‐UTP as a terminator by two low‐fidelity polymerases but not by a high‐fidelity polymerase. The sequence of the primer and template used for these extension reactions is shown at the top of the figure. Polymerase extension reactions were performed by incubating the primer and template with 2’‐F,Me‐UTP and the appropriate reaction buffer for the specific enzyme, followed by detection of the reaction products by MALDI‐TOF MS. The MS spectra of the extension products generated by <t>Therminator</t> II (T2) in (A) and Therminator IX (T9) in (B) indicate single‐base incorporation and termination, whereas the MS spectrum for Thermo Sequenase (TS) in (C) indicates no incorporation, showing only a primer peak. The accuracy for m/z determination is ± 10 Da
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    Template-directed assembly of Z-TFOs. ( A ) Strategy for the assembly of oligonucleotides containing a triplex-forming sequence (TFS; shown in red) containing Z. (i) Primer extension of a template that contains the W–C complement to the TFO (underlined) allows the incorporation of Z by the formation of a GZ base pair at high pH (and the absence of dCTP). (ii) The modified oligonucleotide can then be isolated by selective degradation of the phosphate-labelled template (in yellow) by the action of lambda exonuclease. ( B ) Electrophoretic mobility shift assay showing the products of the extension and digestion of such reactions. The composition of each sample is shown above each lane of the gel. Final concentration of the strands and dNTPS was 5 and 100 μM, respectively. Extension reactions were performed for 2 h at 72°C with 2 units of Therminator Pol, whilst digestion reactions were performed for 2 h with 10 units of Lambda exonuclease at 37°C. The complexes were then separated on a 15% non-denaturing polyacrylamide gel and subjected to post-staining with GelRed.

    Journal: Nucleic Acids Research

    Article Title: Triplex-forming properties and enzymatic incorporation of a base-modified nucleotide capable of duplex DNA recognition at neutral pH

    doi: 10.1093/nar/gkab572

    Figure Lengend Snippet: Template-directed assembly of Z-TFOs. ( A ) Strategy for the assembly of oligonucleotides containing a triplex-forming sequence (TFS; shown in red) containing Z. (i) Primer extension of a template that contains the W–C complement to the TFO (underlined) allows the incorporation of Z by the formation of a GZ base pair at high pH (and the absence of dCTP). (ii) The modified oligonucleotide can then be isolated by selective degradation of the phosphate-labelled template (in yellow) by the action of lambda exonuclease. ( B ) Electrophoretic mobility shift assay showing the products of the extension and digestion of such reactions. The composition of each sample is shown above each lane of the gel. Final concentration of the strands and dNTPS was 5 and 100 μM, respectively. Extension reactions were performed for 2 h at 72°C with 2 units of Therminator Pol, whilst digestion reactions were performed for 2 h with 10 units of Lambda exonuclease at 37°C. The complexes were then separated on a 15% non-denaturing polyacrylamide gel and subjected to post-staining with GelRed.

    Article Snippet: The reaction mixture was pre-incubated at 72°C for 30 s before addition of 2 units of Therminator® polymerase (New England Biolabs).

    Techniques: Sequencing, Modification, Isolation, Electrophoretic Mobility Shift Assay, Concentration Assay, Staining

    PAGE analysis of the incorporation of PEG modified dUTPs with the use of Vent ( exo- ), Therminator, Pfu and Bst 2.0 DNA polymerases. All lanes except “no dNTPs” contain dATP, dCTP, dGTP and the nucleotide indicated above the gel (from left): Lane 1 = dTTP, dATP, dCTP, dGTP; Lane 2 = PEG 8 -dUTP ( 22 ), dATP, dCTP, dGTP; Lane 3 = PEG 16 -dUTP ( 23 ), dATP, dCTP, dGTP; Lane 4 = PEG 24 -dUTP ( 24 ), dATP, dCTP, dGTP; Lane 5 = PEG 6 -dUTP, dATP, dCTP, dGTP; Lane 6 = dATP, dCTP, dGTP; Lane 7 = Control, no dNTPs present. Conditions for all primer reactions; 20 nM primer, 15 nM template, 625 nM dNTPs, 40 °C, 1 h.

    Journal: Molecules

    Article Title: Synthesis of Polyanionic C5-Modified 2′-Deoxyuridine and 2′-Deoxycytidine-5′-Triphosphates and Their Properties as Substrates for DNA Polymerases

    doi: 10.3390/molecules26082250

    Figure Lengend Snippet: PAGE analysis of the incorporation of PEG modified dUTPs with the use of Vent ( exo- ), Therminator, Pfu and Bst 2.0 DNA polymerases. All lanes except “no dNTPs” contain dATP, dCTP, dGTP and the nucleotide indicated above the gel (from left): Lane 1 = dTTP, dATP, dCTP, dGTP; Lane 2 = PEG 8 -dUTP ( 22 ), dATP, dCTP, dGTP; Lane 3 = PEG 16 -dUTP ( 23 ), dATP, dCTP, dGTP; Lane 4 = PEG 24 -dUTP ( 24 ), dATP, dCTP, dGTP; Lane 5 = PEG 6 -dUTP, dATP, dCTP, dGTP; Lane 6 = dATP, dCTP, dGTP; Lane 7 = Control, no dNTPs present. Conditions for all primer reactions; 20 nM primer, 15 nM template, 625 nM dNTPs, 40 °C, 1 h.

    Article Snippet: Using either ploymerases HotStarTaq (heated to 95 °C for 3 min prior to use), Phusion, Vent ( exo- ), Klenow ( exo- ), Bst 2.0, Therminator (New England Biolabs, Ipswich, MA, USA), or Omni-KlenTaq DNA polymerase (DNA Technology, St Louis, MO, USA), primer extension reactions were performed as 20 µL reactions comprising of: 1 U polymerase in 1× polymerase specific buffer (provided by DNA Technology, St Louis, MO, USA), 20 nM Cy5-5′-labelled primer and 15 nM template.

    Techniques: Modification

    PAGE analysis of the incorporation of PEG modified dCTPs with the use of Omni-Klentaq, Therminator, Klenow ( exo- ) and Vent ( exo- ) DNA polymerase. All lanes except “no dNTPs” contain dATP, dGTP, dTTP and the nucleotide indicated above the gel (from left): Lane 1 = dCTP, dATP, dGTP, dTTP; Lane 2 = PEG 8 -dCTP ( 25 ), dATP, dGTP, dTTP; Lane 3 = PEG 4 -dCTP ( 26 ), dATP, dGTP, dTTP; Lane 4 = dATP, dGTP, dTTP; Lane 5 = Control, no dNTPs present. Conditions for all primer reactions; 20 nM primer, 15 nM template, 625 nM dNTPs, 40 °C, 1 h.

    Journal: Molecules

    Article Title: Synthesis of Polyanionic C5-Modified 2′-Deoxyuridine and 2′-Deoxycytidine-5′-Triphosphates and Their Properties as Substrates for DNA Polymerases

    doi: 10.3390/molecules26082250

    Figure Lengend Snippet: PAGE analysis of the incorporation of PEG modified dCTPs with the use of Omni-Klentaq, Therminator, Klenow ( exo- ) and Vent ( exo- ) DNA polymerase. All lanes except “no dNTPs” contain dATP, dGTP, dTTP and the nucleotide indicated above the gel (from left): Lane 1 = dCTP, dATP, dGTP, dTTP; Lane 2 = PEG 8 -dCTP ( 25 ), dATP, dGTP, dTTP; Lane 3 = PEG 4 -dCTP ( 26 ), dATP, dGTP, dTTP; Lane 4 = dATP, dGTP, dTTP; Lane 5 = Control, no dNTPs present. Conditions for all primer reactions; 20 nM primer, 15 nM template, 625 nM dNTPs, 40 °C, 1 h.

    Article Snippet: Using either ploymerases HotStarTaq (heated to 95 °C for 3 min prior to use), Phusion, Vent ( exo- ), Klenow ( exo- ), Bst 2.0, Therminator (New England Biolabs, Ipswich, MA, USA), or Omni-KlenTaq DNA polymerase (DNA Technology, St Louis, MO, USA), primer extension reactions were performed as 20 µL reactions comprising of: 1 U polymerase in 1× polymerase specific buffer (provided by DNA Technology, St Louis, MO, USA), 20 nM Cy5-5′-labelled primer and 15 nM template.

    Techniques: Modification

    Incorporation of 2’‐F,Me‐UTP as a terminator by two low‐fidelity polymerases but not by a high‐fidelity polymerase. The sequence of the primer and template used for these extension reactions is shown at the top of the figure. Polymerase extension reactions were performed by incubating the primer and template with 2’‐F,Me‐UTP and the appropriate reaction buffer for the specific enzyme, followed by detection of the reaction products by MALDI‐TOF MS. The MS spectra of the extension products generated by Therminator II (T2) in (A) and Therminator IX (T9) in (B) indicate single‐base incorporation and termination, whereas the MS spectrum for Thermo Sequenase (TS) in (C) indicates no incorporation, showing only a primer peak. The accuracy for m/z determination is ± 10 Da

    Journal: Pharmacology Research & Perspectives

    Article Title: Nucleotide analogues as inhibitors of SARS‐CoV Polymerase

    doi: 10.1002/prp2.674

    Figure Lengend Snippet: Incorporation of 2’‐F,Me‐UTP as a terminator by two low‐fidelity polymerases but not by a high‐fidelity polymerase. The sequence of the primer and template used for these extension reactions is shown at the top of the figure. Polymerase extension reactions were performed by incubating the primer and template with 2’‐F,Me‐UTP and the appropriate reaction buffer for the specific enzyme, followed by detection of the reaction products by MALDI‐TOF MS. The MS spectra of the extension products generated by Therminator II (T2) in (A) and Therminator IX (T9) in (B) indicate single‐base incorporation and termination, whereas the MS spectrum for Thermo Sequenase (TS) in (C) indicates no incorporation, showing only a primer peak. The accuracy for m/z determination is ± 10 Da

    Article Snippet: The 20 µl extension reactions consisted of 3 µM DNA template and 5 µM DNA primer (sequences shown in Fig. ), 10 µM 2’‐F,Me‐UTP (Sierra Bioresearch), 1x Thermo Sequenase buffer or 1x ThermoPol buffer (for Therminator enzymes), and either 10 U Thermo Sequenase (GE Healthcare), 4 U Therminator II or 10 U Therminator IX (New England Biolabs).

    Techniques: Sequencing, Generated