cohesive end taq ligase  (New England Biolabs)


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    Structured Review

    New England Biolabs cohesive end taq ligase
    Cohesive End Taq Ligase, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cohesive end taq ligase/product/New England Biolabs
    Average 96 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    cohesive end taq ligase - by Bioz Stars, 2020-03
    96/100 stars

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    Centrifugation:

    Article Title: Engineering selective competitors for the discrimination of highly conserved protein-protein interaction modules
    Article Snippet: For the elution step, 100 µL of buffer EB (Qiagen kit) was added to the column and incubated for 10 min. After a last centrifugation of 30 s, the eluant containing the purified dU-ssDNA was saved. .. The annealing was performed by heating to 95 °C for 3 min, then at 60 °C for 3 min. After the annealing step, 200 units of cohesive end Taq ligase (New England BioLabs) and 2.5 units of PfuTurbo Cx hotstart DNA polymerase were added bringing the total volume to 100 µL.

    Ligation:

    Article Title: Engineering selective competitors for the discrimination of highly conserved protein-protein interaction modules
    Article Snippet: The annealing was performed by heating to 95 °C for 3 min, then at 60 °C for 3 min. After the annealing step, 200 units of cohesive end Taq ligase (New England BioLabs) and 2.5 units of PfuTurbo Cx hotstart DNA polymerase were added bringing the total volume to 100 µL. .. Extension and ligation of the mutant strand was performed at 65 °C for 15 min and 45 °C for 15 min. A total of 3.8 pmol of phosphorylated oligonucleotide (5′-P-ggtctggtgctggcattctc) was added and one more cycle of 95 °C for 30 s, 55 °C for 45 s, 65 °C for 10 min and 45 °C for 15 min was performed.

    Mutagenesis:

    Article Title: Engineering selective competitors for the discrimination of highly conserved protein-protein interaction modules
    Article Snippet: The ssDNA concentration was determined with a Nanodrop spectrophotometer (Thermo Scientific) and the quality was evaluated by electrophoresing 3 µL on a 0.8% TAE/agarose gel at 100 V for 1 h. Combinatorial libraries in which the BC and FG loops of 10 FN3 were diversified were constructed by using pFunkel mutagenesis , inspired by Kunkel mutagenesis. .. The annealing was performed by heating to 95 °C for 3 min, then at 60 °C for 3 min. After the annealing step, 200 units of cohesive end Taq ligase (New England BioLabs) and 2.5 units of PfuTurbo Cx hotstart DNA polymerase were added bringing the total volume to 100 µL.

    Construct:

    Article Title: Engineering selective competitors for the discrimination of highly conserved protein-protein interaction modules
    Article Snippet: The ssDNA concentration was determined with a Nanodrop spectrophotometer (Thermo Scientific) and the quality was evaluated by electrophoresing 3 µL on a 0.8% TAE/agarose gel at 100 V for 1 h. Combinatorial libraries in which the BC and FG loops of 10 FN3 were diversified were constructed by using pFunkel mutagenesis , inspired by Kunkel mutagenesis. .. The annealing was performed by heating to 95 °C for 3 min, then at 60 °C for 3 min. After the annealing step, 200 units of cohesive end Taq ligase (New England BioLabs) and 2.5 units of PfuTurbo Cx hotstart DNA polymerase were added bringing the total volume to 100 µL.

    Purification:

    Article Title: Engineering selective competitors for the discrimination of highly conserved protein-protein interaction modules
    Article Snippet: For the elution step, 100 µL of buffer EB (Qiagen kit) was added to the column and incubated for 10 min. After a last centrifugation of 30 s, the eluant containing the purified dU-ssDNA was saved. .. The annealing was performed by heating to 95 °C for 3 min, then at 60 °C for 3 min. After the annealing step, 200 units of cohesive end Taq ligase (New England BioLabs) and 2.5 units of PfuTurbo Cx hotstart DNA polymerase were added bringing the total volume to 100 µL.

    Concentration Assay:

    Article Title: Engineering selective competitors for the discrimination of highly conserved protein-protein interaction modules
    Article Snippet: The ssDNA concentration was determined with a Nanodrop spectrophotometer (Thermo Scientific) and the quality was evaluated by electrophoresing 3 µL on a 0.8% TAE/agarose gel at 100 V for 1 h. Combinatorial libraries in which the BC and FG loops of 10 FN3 were diversified were constructed by using pFunkel mutagenesis , inspired by Kunkel mutagenesis. .. The annealing was performed by heating to 95 °C for 3 min, then at 60 °C for 3 min. After the annealing step, 200 units of cohesive end Taq ligase (New England BioLabs) and 2.5 units of PfuTurbo Cx hotstart DNA polymerase were added bringing the total volume to 100 µL.

    Incubation:

    Article Title: Engineering selective competitors for the discrimination of highly conserved protein-protein interaction modules
    Article Snippet: For the elution step, 100 µL of buffer EB (Qiagen kit) was added to the column and incubated for 10 min. After a last centrifugation of 30 s, the eluant containing the purified dU-ssDNA was saved. .. The annealing was performed by heating to 95 °C for 3 min, then at 60 °C for 3 min. After the annealing step, 200 units of cohesive end Taq ligase (New England BioLabs) and 2.5 units of PfuTurbo Cx hotstart DNA polymerase were added bringing the total volume to 100 µL.

    Introduce:

    Article Title: Engineering selective competitors for the discrimination of highly conserved protein-protein interaction modules
    Article Snippet: Five phosphorylated oligonucleotides were designed to introduce from 5 to 9 NNK random codons at position 25 in the 10 FN3 gene and seven phosphorylated oligonucleotides were designed to introduce 7 to 13 NNK random codons at position 75 corresponding to the BC and the FG 10 FN3 loops, respectively (see Supplementary Note for the oligonucleotides sequence; the numbering corresponds to the one used for the wild-type 10 FN3, PDB ID 1FNA). .. The annealing was performed by heating to 95 °C for 3 min, then at 60 °C for 3 min. After the annealing step, 200 units of cohesive end Taq ligase (New England BioLabs) and 2.5 units of PfuTurbo Cx hotstart DNA polymerase were added bringing the total volume to 100 µL.

    Sequencing:

    Article Title: Engineering selective competitors for the discrimination of highly conserved protein-protein interaction modules
    Article Snippet: Five phosphorylated oligonucleotides were designed to introduce from 5 to 9 NNK random codons at position 25 in the 10 FN3 gene and seven phosphorylated oligonucleotides were designed to introduce 7 to 13 NNK random codons at position 75 corresponding to the BC and the FG 10 FN3 loops, respectively (see Supplementary Note for the oligonucleotides sequence; the numbering corresponds to the one used for the wild-type 10 FN3, PDB ID 1FNA). .. The annealing was performed by heating to 95 °C for 3 min, then at 60 °C for 3 min. After the annealing step, 200 units of cohesive end Taq ligase (New England BioLabs) and 2.5 units of PfuTurbo Cx hotstart DNA polymerase were added bringing the total volume to 100 µL.

    Transformation Assay:

    Article Title: Engineering selective competitors for the discrimination of highly conserved protein-protein interaction modules
    Article Snippet: The annealing was performed by heating to 95 °C for 3 min, then at 60 °C for 3 min. After the annealing step, 200 units of cohesive end Taq ligase (New England BioLabs) and 2.5 units of PfuTurbo Cx hotstart DNA polymerase were added bringing the total volume to 100 µL. .. Before purification, 1 µL of the pFunkel solution was transformed into 10 µL of E. cloni 10G chemically competent cells (Lucigen, 60106-2) and were plated onto 2x YT medium agar-plate with 50 µg mL−1 carbenicillin, 10 mM glucose and incubated at 37 °C overnight.

    Spectrophotometry:

    Article Title: Engineering selective competitors for the discrimination of highly conserved protein-protein interaction modules
    Article Snippet: The ssDNA concentration was determined with a Nanodrop spectrophotometer (Thermo Scientific) and the quality was evaluated by electrophoresing 3 µL on a 0.8% TAE/agarose gel at 100 V for 1 h. Combinatorial libraries in which the BC and FG loops of 10 FN3 were diversified were constructed by using pFunkel mutagenesis , inspired by Kunkel mutagenesis. .. The annealing was performed by heating to 95 °C for 3 min, then at 60 °C for 3 min. After the annealing step, 200 units of cohesive end Taq ligase (New England BioLabs) and 2.5 units of PfuTurbo Cx hotstart DNA polymerase were added bringing the total volume to 100 µL.

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  • 96
    New England Biolabs cohesive end taq ligase
    Cohesive End Taq Ligase, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/cohesive end taq ligase/product/New England Biolabs
    Average 96 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    cohesive end taq ligase - by Bioz Stars, 2020-03
    96/100 stars
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