t4 dna ligase Search Results


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  • 78
    Thermo Fisher t4 dna ligase ligated plasmids
    T4 Dna Ligase Ligated Plasmids, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 78/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    77
    New England Biolabs t4 dna ligase ligation buffer
    T4 Dna Ligase Ligation Buffer, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 77/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    98
    TaKaRa t4 dna ligase
    Dependence of the efficiency of DNA ligation using <t>T4</t> DNA ligase immobilized on ferromagnetic particles in the absence of a magnetic field on the ambient temperature. The ordinate axis represents the ligation efficiency, which is normalized by that at 16 °C. The standard deviations are obtained from 6 independent experiments.
    T4 Dna Ligase, supplied by TaKaRa, used in various techniques. Bioz Stars score: 98/100, based on 5252 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    70
    New England Biolabs ligation t4 dna ligase
    Dependence of the efficiency of DNA ligation using <t>T4</t> DNA ligase immobilized on ferromagnetic particles in the absence of a magnetic field on the ambient temperature. The ordinate axis represents the ligation efficiency, which is normalized by that at 16 °C. The standard deviations are obtained from 6 independent experiments.
    Ligation T4 Dna Ligase, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 70/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher t4 dna ligase buffer
    Dependence of the efficiency of DNA ligation using <t>T4</t> DNA ligase immobilized on ferromagnetic particles in the absence of a magnetic field on the ambient temperature. The ordinate axis represents the ligation efficiency, which is normalized by that at 16 °C. The standard deviations are obtained from 6 independent experiments.
    T4 Dna Ligase Buffer, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 541 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Roche t4 dna ligase
    Dependence of the efficiency of DNA ligation using <t>T4</t> DNA ligase immobilized on ferromagnetic particles in the absence of a magnetic field on the ambient temperature. The ordinate axis represents the ligation efficiency, which is normalized by that at 16 °C. The standard deviations are obtained from 6 independent experiments.
    T4 Dna Ligase, supplied by Roche, used in various techniques. Bioz Stars score: 97/100, based on 3717 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Promega t4 dna ligase
    Partial stimulation of uracil repair by the addition of exogenous proteins. Duplex DNA containing U opposite G at a Hpa II site was incubated at 30°C for 3 h with Arabidopsis cell extract in a reaction mixture containing either deoxycytidine triphosphate (dCTP) or all four deoxyribonucleotide triphosphates (dNTPs), as indicated, and different combinations of <t>T4</t> DNA ligase (1.5 U), Arabidopsis AtXRCC1 (65 n m ) and human Pol β (2.4 U). Reaction products were digested with Hpa II, separated in a 12% denaturing polyacrylamide gel and quantified by fluorescence scanning. The percentage of fully repaired DNA product is shown. Values are means with standard errors from two independent experiments.
    T4 Dna Ligase, supplied by Promega, used in various techniques. Bioz Stars score: 98/100, based on 8221 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher t4 dna ligase
    Partial stimulation of uracil repair by the addition of exogenous proteins. Duplex DNA containing U opposite G at a Hpa II site was incubated at 30°C for 3 h with Arabidopsis cell extract in a reaction mixture containing either deoxycytidine triphosphate (dCTP) or all four deoxyribonucleotide triphosphates (dNTPs), as indicated, and different combinations of <t>T4</t> DNA ligase (1.5 U), Arabidopsis AtXRCC1 (65 n m ) and human Pol β (2.4 U). Reaction products were digested with Hpa II, separated in a 12% denaturing polyacrylamide gel and quantified by fluorescence scanning. The percentage of fully repaired DNA product is shown. Values are means with standard errors from two independent experiments.
    T4 Dna Ligase, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 18366 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Toyobo t4 dna ligase
    Partial stimulation of uracil repair by the addition of exogenous proteins. Duplex DNA containing U opposite G at a Hpa II site was incubated at 30°C for 3 h with Arabidopsis cell extract in a reaction mixture containing either deoxycytidine triphosphate (dCTP) or all four deoxyribonucleotide triphosphates (dNTPs), as indicated, and different combinations of <t>T4</t> DNA ligase (1.5 U), Arabidopsis AtXRCC1 (65 n m ) and human Pol β (2.4 U). Reaction products were digested with Hpa II, separated in a 12% denaturing polyacrylamide gel and quantified by fluorescence scanning. The percentage of fully repaired DNA product is shown. Values are means with standard errors from two independent experiments.
    T4 Dna Ligase, supplied by Toyobo, used in various techniques. Bioz Stars score: 97/100, based on 245 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Enzymatics t4 dna ligase
    Partial stimulation of uracil repair by the addition of exogenous proteins. Duplex DNA containing U opposite G at a Hpa II site was incubated at 30°C for 3 h with Arabidopsis cell extract in a reaction mixture containing either deoxycytidine triphosphate (dCTP) or all four deoxyribonucleotide triphosphates (dNTPs), as indicated, and different combinations of <t>T4</t> DNA ligase (1.5 U), Arabidopsis AtXRCC1 (65 n m ) and human Pol β (2.4 U). Reaction products were digested with Hpa II, separated in a 12% denaturing polyacrylamide gel and quantified by fluorescence scanning. The percentage of fully repaired DNA product is shown. Values are means with standard errors from two independent experiments.
    T4 Dna Ligase, supplied by Enzymatics, used in various techniques. Bioz Stars score: 97/100, based on 636 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    GE Healthcare t4 dna ligase
    Partial stimulation of uracil repair by the addition of exogenous proteins. Duplex DNA containing U opposite G at a Hpa II site was incubated at 30°C for 3 h with Arabidopsis cell extract in a reaction mixture containing either deoxycytidine triphosphate (dCTP) or all four deoxyribonucleotide triphosphates (dNTPs), as indicated, and different combinations of <t>T4</t> DNA ligase (1.5 U), Arabidopsis AtXRCC1 (65 n m ) and human Pol β (2.4 U). Reaction products were digested with Hpa II, separated in a 12% denaturing polyacrylamide gel and quantified by fluorescence scanning. The percentage of fully repaired DNA product is shown. Values are means with standard errors from two independent experiments.
    T4 Dna Ligase, supplied by GE Healthcare, used in various techniques. Bioz Stars score: 93/100, based on 459 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Shanghai Genechem t4 dna ligase
    Partial stimulation of uracil repair by the addition of exogenous proteins. Duplex DNA containing U opposite G at a Hpa II site was incubated at 30°C for 3 h with Arabidopsis cell extract in a reaction mixture containing either deoxycytidine triphosphate (dCTP) or all four deoxyribonucleotide triphosphates (dNTPs), as indicated, and different combinations of <t>T4</t> DNA ligase (1.5 U), Arabidopsis AtXRCC1 (65 n m ) and human Pol β (2.4 U). Reaction products were digested with Hpa II, separated in a 12% denaturing polyacrylamide gel and quantified by fluorescence scanning. The percentage of fully repaired DNA product is shown. Values are means with standard errors from two independent experiments.
    T4 Dna Ligase, supplied by Shanghai Genechem, used in various techniques. Bioz Stars score: 97/100, based on 21 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Fisher Scientific t4 dna ligase
    Partial stimulation of uracil repair by the addition of exogenous proteins. Duplex DNA containing U opposite G at a Hpa II site was incubated at 30°C for 3 h with Arabidopsis cell extract in a reaction mixture containing either deoxycytidine triphosphate (dCTP) or all four deoxyribonucleotide triphosphates (dNTPs), as indicated, and different combinations of <t>T4</t> DNA ligase (1.5 U), Arabidopsis AtXRCC1 (65 n m ) and human Pol β (2.4 U). Reaction products were digested with Hpa II, separated in a 12% denaturing polyacrylamide gel and quantified by fluorescence scanning. The percentage of fully repaired DNA product is shown. Values are means with standard errors from two independent experiments.
    T4 Dna Ligase, supplied by Fisher Scientific, used in various techniques. Bioz Stars score: 94/100, based on 141 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Bio-Rad t4 dna ligase
    Partial stimulation of uracil repair by the addition of exogenous proteins. Duplex DNA containing U opposite G at a Hpa II site was incubated at 30°C for 3 h with Arabidopsis cell extract in a reaction mixture containing either deoxycytidine triphosphate (dCTP) or all four deoxyribonucleotide triphosphates (dNTPs), as indicated, and different combinations of <t>T4</t> DNA ligase (1.5 U), Arabidopsis AtXRCC1 (65 n m ) and human Pol β (2.4 U). Reaction products were digested with Hpa II, separated in a 12% denaturing polyacrylamide gel and quantified by fluorescence scanning. The percentage of fully repaired DNA product is shown. Values are means with standard errors from two independent experiments.
    T4 Dna Ligase, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 97/100, based on 80 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore t4 dna ligase
    Partial stimulation of uracil repair by the addition of exogenous proteins. Duplex DNA containing U opposite G at a Hpa II site was incubated at 30°C for 3 h with Arabidopsis cell extract in a reaction mixture containing either deoxycytidine triphosphate (dCTP) or all four deoxyribonucleotide triphosphates (dNTPs), as indicated, and different combinations of <t>T4</t> DNA ligase (1.5 U), Arabidopsis AtXRCC1 (65 n m ) and human Pol β (2.4 U). Reaction products were digested with Hpa II, separated in a 12% denaturing polyacrylamide gel and quantified by fluorescence scanning. The percentage of fully repaired DNA product is shown. Values are means with standard errors from two independent experiments.
    T4 Dna Ligase, supplied by Millipore, used in various techniques. Bioz Stars score: 97/100, based on 468 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Boehringer Mannheim t4 dna ligase
    Partial stimulation of uracil repair by the addition of exogenous proteins. Duplex DNA containing U opposite G at a Hpa II site was incubated at 30°C for 3 h with Arabidopsis cell extract in a reaction mixture containing either deoxycytidine triphosphate (dCTP) or all four deoxyribonucleotide triphosphates (dNTPs), as indicated, and different combinations of <t>T4</t> DNA ligase (1.5 U), Arabidopsis AtXRCC1 (65 n m ) and human Pol β (2.4 U). Reaction products were digested with Hpa II, separated in a 12% denaturing polyacrylamide gel and quantified by fluorescence scanning. The percentage of fully repaired DNA product is shown. Values are means with standard errors from two independent experiments.
    T4 Dna Ligase, supplied by Boehringer Mannheim, used in various techniques. Bioz Stars score: 93/100, based on 855 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Beijing CWBio t4 dna ligase
    Partial stimulation of uracil repair by the addition of exogenous proteins. Duplex DNA containing U opposite G at a Hpa II site was incubated at 30°C for 3 h with Arabidopsis cell extract in a reaction mixture containing either deoxycytidine triphosphate (dCTP) or all four deoxyribonucleotide triphosphates (dNTPs), as indicated, and different combinations of <t>T4</t> DNA ligase (1.5 U), Arabidopsis AtXRCC1 (65 n m ) and human Pol β (2.4 U). Reaction products were digested with Hpa II, separated in a 12% denaturing polyacrylamide gel and quantified by fluorescence scanning. The percentage of fully repaired DNA product is shown. Values are means with standard errors from two independent experiments.
    T4 Dna Ligase, supplied by Beijing CWBio, used in various techniques. Bioz Stars score: 93/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Agilent technologies t4 dna ligase
    Partial stimulation of uracil repair by the addition of exogenous proteins. Duplex DNA containing U opposite G at a Hpa II site was incubated at 30°C for 3 h with Arabidopsis cell extract in a reaction mixture containing either deoxycytidine triphosphate (dCTP) or all four deoxyribonucleotide triphosphates (dNTPs), as indicated, and different combinations of <t>T4</t> DNA ligase (1.5 U), Arabidopsis AtXRCC1 (65 n m ) and human Pol β (2.4 U). Reaction products were digested with Hpa II, separated in a 12% denaturing polyacrylamide gel and quantified by fluorescence scanning. The percentage of fully repaired DNA product is shown. Values are means with standard errors from two independent experiments.
    T4 Dna Ligase, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 97/100, based on 156 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    FUJIFILM t4 dna ligase
    Partial stimulation of uracil repair by the addition of exogenous proteins. Duplex DNA containing U opposite G at a Hpa II site was incubated at 30°C for 3 h with Arabidopsis cell extract in a reaction mixture containing either deoxycytidine triphosphate (dCTP) or all four deoxyribonucleotide triphosphates (dNTPs), as indicated, and different combinations of <t>T4</t> DNA ligase (1.5 U), Arabidopsis AtXRCC1 (65 n m ) and human Pol β (2.4 U). Reaction products were digested with Hpa II, separated in a 12% denaturing polyacrylamide gel and quantified by fluorescence scanning. The percentage of fully repaired DNA product is shown. Values are means with standard errors from two independent experiments.
    T4 Dna Ligase, supplied by FUJIFILM, used in various techniques. Bioz Stars score: 93/100, based on 34 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    tiangen biotech co t4 dna ligase
    Partial stimulation of uracil repair by the addition of exogenous proteins. Duplex DNA containing U opposite G at a Hpa II site was incubated at 30°C for 3 h with Arabidopsis cell extract in a reaction mixture containing either deoxycytidine triphosphate (dCTP) or all four deoxyribonucleotide triphosphates (dNTPs), as indicated, and different combinations of <t>T4</t> DNA ligase (1.5 U), Arabidopsis AtXRCC1 (65 n m ) and human Pol β (2.4 U). Reaction products were digested with Hpa II, separated in a 12% denaturing polyacrylamide gel and quantified by fluorescence scanning. The percentage of fully repaired DNA product is shown. Values are means with standard errors from two independent experiments.
    T4 Dna Ligase, supplied by tiangen biotech co, used in various techniques. Bioz Stars score: 97/100, based on 64 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    t4 dna ligase - by Bioz Stars, 2020-01
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    Monserate Biotechnology Group t4 dna ligase
    Partial stimulation of uracil repair by the addition of exogenous proteins. Duplex DNA containing U opposite G at a Hpa II site was incubated at 30°C for 3 h with Arabidopsis cell extract in a reaction mixture containing either deoxycytidine triphosphate (dCTP) or all four deoxyribonucleotide triphosphates (dNTPs), as indicated, and different combinations of <t>T4</t> DNA ligase (1.5 U), Arabidopsis AtXRCC1 (65 n m ) and human Pol β (2.4 U). Reaction products were digested with Hpa II, separated in a 12% denaturing polyacrylamide gel and quantified by fluorescence scanning. The percentage of fully repaired DNA product is shown. Values are means with standard errors from two independent experiments.
    T4 Dna Ligase, supplied by Monserate Biotechnology Group, used in various techniques. Bioz Stars score: 93/100, based on 38 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    PerkinElmer t4 dna ligase
    Partial stimulation of uracil repair by the addition of exogenous proteins. Duplex DNA containing U opposite G at a Hpa II site was incubated at 30°C for 3 h with Arabidopsis cell extract in a reaction mixture containing either deoxycytidine triphosphate (dCTP) or all four deoxyribonucleotide triphosphates (dNTPs), as indicated, and different combinations of <t>T4</t> DNA ligase (1.5 U), Arabidopsis AtXRCC1 (65 n m ) and human Pol β (2.4 U). Reaction products were digested with Hpa II, separated in a 12% denaturing polyacrylamide gel and quantified by fluorescence scanning. The percentage of fully repaired DNA product is shown. Values are means with standard errors from two independent experiments.
    T4 Dna Ligase, supplied by PerkinElmer, used in various techniques. Bioz Stars score: 93/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Nanocs t4 dna ligase
    Optimization of D-probe complementary sequence length. (A) Complementary sequences of four probe sets for miR-143. (B) The fluorescence intensities of D-probes bound to C-probes in the presence of <t>T4</t> DNA ligase. Data represent the mean ± S.E. (n = 3). (C) Correlation between input miR-143 and the signal of D-probe-143-(8) in the presence (filled circle) or absence (open circle) of T4 DNA ligase. The upper axis indicates the final concentration of input miR-143 in the hybridization chamber while the lower axis indicates the total amount of input miR-143. Data in the linear range were fitted by a linear expression (gray lines). Data represent the mean ± S.E. (n = 3).
    T4 Dna Ligase, supplied by Nanocs, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Kapa Biosystems t4 dna ligase
    Optimization of D-probe complementary sequence length. (A) Complementary sequences of four probe sets for miR-143. (B) The fluorescence intensities of D-probes bound to C-probes in the presence of <t>T4</t> DNA ligase. Data represent the mean ± S.E. (n = 3). (C) Correlation between input miR-143 and the signal of D-probe-143-(8) in the presence (filled circle) or absence (open circle) of T4 DNA ligase. The upper axis indicates the final concentration of input miR-143 in the hybridization chamber while the lower axis indicates the total amount of input miR-143. Data in the linear range were fitted by a linear expression (gray lines). Data represent the mean ± S.E. (n = 3).
    T4 Dna Ligase, supplied by Kapa Biosystems, used in various techniques. Bioz Stars score: 97/100, based on 33 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    N/A
    T4 DNA Ligase can catalyze the formation of a phosphodiester bond between juxtaposed 5 phosphate and 3 hydroxyl termini in duplex DNA or RNA This enzyme will join blunt end
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    N/A
    T4 DNA Ligase catalyzes the formation of a phosphodiester bond between juxtaposed 5 phosphate and 3 hydroxyl termini in duplex DNA or RNA
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    Image Search Results


    Dependence of the efficiency of DNA ligation using T4 DNA ligase immobilized on ferromagnetic particles in the absence of a magnetic field on the ambient temperature. The ordinate axis represents the ligation efficiency, which is normalized by that at 16 °C. The standard deviations are obtained from 6 independent experiments.

    Journal: Biochemistry and Biophysics Reports

    Article Title: Efficient DNA ligation by selective heating of DNA ligase with a radio frequency alternating magnetic field

    doi: 10.1016/j.bbrep.2016.10.006

    Figure Lengend Snippet: Dependence of the efficiency of DNA ligation using T4 DNA ligase immobilized on ferromagnetic particles in the absence of a magnetic field on the ambient temperature. The ordinate axis represents the ligation efficiency, which is normalized by that at 16 °C. The standard deviations are obtained from 6 independent experiments.

    Article Snippet: In summary, we carried out the ligation of DNA fragments with cohesive ends using T4 DNA ligase immobilized on ferromagnetic particles and found that the ligation efficiency was increased under a radio frequency alternating magnetic field caused by heat generation from the particles.

    Techniques: DNA Ligation, Ligation

    Dependence of the efficiency of DNA ligation using T4 DNA ligase immobilized on ferromagnetic particles under an ac magnetic field of 0.34 MHz on the amplitude of the magnetic field. The ambient temperature is 16 °C. The ordinate axis represents the ligation efficiency under an ac magnetic field, which is normalized by that in the absence of a magnetic field. The inset shows the ligation efficiency under the ac magnetic field as a function of the average surface temperature of ferromagnetic particles, noting that the surface temperature increases with an increase in the field amplitude. The standard deviations are obtained from 6 independent experiments.

    Journal: Biochemistry and Biophysics Reports

    Article Title: Efficient DNA ligation by selective heating of DNA ligase with a radio frequency alternating magnetic field

    doi: 10.1016/j.bbrep.2016.10.006

    Figure Lengend Snippet: Dependence of the efficiency of DNA ligation using T4 DNA ligase immobilized on ferromagnetic particles under an ac magnetic field of 0.34 MHz on the amplitude of the magnetic field. The ambient temperature is 16 °C. The ordinate axis represents the ligation efficiency under an ac magnetic field, which is normalized by that in the absence of a magnetic field. The inset shows the ligation efficiency under the ac magnetic field as a function of the average surface temperature of ferromagnetic particles, noting that the surface temperature increases with an increase in the field amplitude. The standard deviations are obtained from 6 independent experiments.

    Article Snippet: In summary, we carried out the ligation of DNA fragments with cohesive ends using T4 DNA ligase immobilized on ferromagnetic particles and found that the ligation efficiency was increased under a radio frequency alternating magnetic field caused by heat generation from the particles.

    Techniques: DNA Ligation, Ligation

    Partial stimulation of uracil repair by the addition of exogenous proteins. Duplex DNA containing U opposite G at a Hpa II site was incubated at 30°C for 3 h with Arabidopsis cell extract in a reaction mixture containing either deoxycytidine triphosphate (dCTP) or all four deoxyribonucleotide triphosphates (dNTPs), as indicated, and different combinations of T4 DNA ligase (1.5 U), Arabidopsis AtXRCC1 (65 n m ) and human Pol β (2.4 U). Reaction products were digested with Hpa II, separated in a 12% denaturing polyacrylamide gel and quantified by fluorescence scanning. The percentage of fully repaired DNA product is shown. Values are means with standard errors from two independent experiments.

    Journal: The Plant Journal

    Article Title: Single-nucleotide and long-patch base excision repair of DNA damage in plants

    doi: 10.1111/j.1365-313X.2009.03994.x

    Figure Lengend Snippet: Partial stimulation of uracil repair by the addition of exogenous proteins. Duplex DNA containing U opposite G at a Hpa II site was incubated at 30°C for 3 h with Arabidopsis cell extract in a reaction mixture containing either deoxycytidine triphosphate (dCTP) or all four deoxyribonucleotide triphosphates (dNTPs), as indicated, and different combinations of T4 DNA ligase (1.5 U), Arabidopsis AtXRCC1 (65 n m ) and human Pol β (2.4 U). Reaction products were digested with Hpa II, separated in a 12% denaturing polyacrylamide gel and quantified by fluorescence scanning. The percentage of fully repaired DNA product is shown. Values are means with standard errors from two independent experiments.

    Article Snippet: E. coli T4 DNA ligase was purchased from Promega ( http://www.promega.com ) and Hpa II was purchased from Roche ( http://www.roche-applied-science.com ).

    Techniques: Incubation, Fluorescence

    Optimization of D-probe complementary sequence length. (A) Complementary sequences of four probe sets for miR-143. (B) The fluorescence intensities of D-probes bound to C-probes in the presence of T4 DNA ligase. Data represent the mean ± S.E. (n = 3). (C) Correlation between input miR-143 and the signal of D-probe-143-(8) in the presence (filled circle) or absence (open circle) of T4 DNA ligase. The upper axis indicates the final concentration of input miR-143 in the hybridization chamber while the lower axis indicates the total amount of input miR-143. Data in the linear range were fitted by a linear expression (gray lines). Data represent the mean ± S.E. (n = 3).

    Journal: PLoS ONE

    Article Title: Label-Free Quantification of MicroRNAs Using Ligase-Assisted Sandwich Hybridization on a DNA Microarray

    doi: 10.1371/journal.pone.0090920

    Figure Lengend Snippet: Optimization of D-probe complementary sequence length. (A) Complementary sequences of four probe sets for miR-143. (B) The fluorescence intensities of D-probes bound to C-probes in the presence of T4 DNA ligase. Data represent the mean ± S.E. (n = 3). (C) Correlation between input miR-143 and the signal of D-probe-143-(8) in the presence (filled circle) or absence (open circle) of T4 DNA ligase. The upper axis indicates the final concentration of input miR-143 in the hybridization chamber while the lower axis indicates the total amount of input miR-143. Data in the linear range were fitted by a linear expression (gray lines). Data represent the mean ± S.E. (n = 3).

    Article Snippet: Silane-PEG-Maleimide (MW 5000), betaine solution, PEG 6000 solution, bovine serum albumin (BSA) and T4 DNA ligase were purchased from NANOCS Inc. (Boston, MA, USA), WAKO (Osaka, Japan), Hampton Research (Aliso Viejo, CA, USA), Sigma Aldrich Japan (Tokyo, Japan) and TAKARA (Shiga, Japan), respectively.

    Techniques: Sequencing, Fluorescence, Concentration Assay, Hybridization, Expressing

    Experimental design of the LASH assay. (A) Schematic representation of the ligase-assisted sandwich hybridization assay (LASH assay) for detection of non-labeled miRNA. A target miRNA hybridizes to complimentary D-probe and C-probe, resulting in a tertiary complex. Both C-probe and D-probe are capped with a phosphate group at the 5′ end and a hydroxyl group at the 3′ end; this permits T4 DNA ligase to ligate the 5′ end of C-probe to the 3′ end of D-probe, and to ligate the 5′ end of D-probe to the 3′ end of the target miRNA. (B) A bright-field image of droplets spotted on a coverslip by an ink-jet machine. Scale bar, 400 µm. (C) Fluorescence images of 6-FAM-labeled C-probe (left) immobilized on a substrate and Alexa647-labeled D-probe (right) bound to C-probe in the presence of 1.25 nM miR-143. Scale bar, 100 µm.

    Journal: PLoS ONE

    Article Title: Label-Free Quantification of MicroRNAs Using Ligase-Assisted Sandwich Hybridization on a DNA Microarray

    doi: 10.1371/journal.pone.0090920

    Figure Lengend Snippet: Experimental design of the LASH assay. (A) Schematic representation of the ligase-assisted sandwich hybridization assay (LASH assay) for detection of non-labeled miRNA. A target miRNA hybridizes to complimentary D-probe and C-probe, resulting in a tertiary complex. Both C-probe and D-probe are capped with a phosphate group at the 5′ end and a hydroxyl group at the 3′ end; this permits T4 DNA ligase to ligate the 5′ end of C-probe to the 3′ end of D-probe, and to ligate the 5′ end of D-probe to the 3′ end of the target miRNA. (B) A bright-field image of droplets spotted on a coverslip by an ink-jet machine. Scale bar, 400 µm. (C) Fluorescence images of 6-FAM-labeled C-probe (left) immobilized on a substrate and Alexa647-labeled D-probe (right) bound to C-probe in the presence of 1.25 nM miR-143. Scale bar, 100 µm.

    Article Snippet: Silane-PEG-Maleimide (MW 5000), betaine solution, PEG 6000 solution, bovine serum albumin (BSA) and T4 DNA ligase were purchased from NANOCS Inc. (Boston, MA, USA), WAKO (Osaka, Japan), Hampton Research (Aliso Viejo, CA, USA), Sigma Aldrich Japan (Tokyo, Japan) and TAKARA (Shiga, Japan), respectively.

    Techniques: Hybridization, Labeling, Fluorescence