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neb 10x kld  (New England Biolabs)


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    Structured Review

    New England Biolabs neb 10x kld
    Neb 10x Kld, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 6159 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/neb+10x+kld/DpnI/pm41986324-190-31-31
    Average 99 stars, based on 6159 article reviews
    neb 10x kld - by Bioz Stars, 2026-10
    99/100 stars

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    Related Articles

    Amplification:

    Article Title: Acclimation of Photosynthesis to the Environment 1 acts as a copper-binding superoxide detoxification enzyme.
    Article Snippet: Briefly, pLIC03-APE1 was amplified via PCR using NEB Q5 High-Fidelity 2X Master Mix in 25 μl reaction volumes containing 10 ng DNA template and 0.5 μM of specific primers pairs carrying the new codon (C128S mutation: TGC-AGC; H163A: CAC-GCG; C247S: TGC-AGC); the PCR protocol was set according to the manufacturer's instructions (amplification was done for 25 cycles with 180 s elongation time each) and annealing temperatures were calculated with the online NEB Tm calculator tool. .. After amplification, to remove the initial DNA template (i.e., without the added mutation) and ligate the PCR products, 1 μl of PCR product was incubated at RT for 10 min with NEB 10X KLD (Kinase, Ligase, DpnI) Enzyme Mix in a 10 μl reaction volume before being used to transform NEB 5-alpha competent E. coli cells, for plasmid isolation and Sanger sequencing (Eurofins Genomics). .. Plasmids with the correct sequence were used to transform Rosetta (DE3)pLysS Competent E. coli Cells (Novagen) for recombinant protein production.

    Mutagenesis:

    Article Title: Acclimation of Photosynthesis to the Environment 1 acts as a copper-binding superoxide detoxification enzyme.
    Article Snippet: Briefly, pLIC03-APE1 was amplified via PCR using NEB Q5 High-Fidelity 2X Master Mix in 25 μl reaction volumes containing 10 ng DNA template and 0.5 μM of specific primers pairs carrying the new codon (C128S mutation: TGC-AGC; H163A: CAC-GCG; C247S: TGC-AGC); the PCR protocol was set according to the manufacturer's instructions (amplification was done for 25 cycles with 180 s elongation time each) and annealing temperatures were calculated with the online NEB Tm calculator tool. .. After amplification, to remove the initial DNA template (i.e., without the added mutation) and ligate the PCR products, 1 μl of PCR product was incubated at RT for 10 min with NEB 10X KLD (Kinase, Ligase, DpnI) Enzyme Mix in a 10 μl reaction volume before being used to transform NEB 5-alpha competent E. coli cells, for plasmid isolation and Sanger sequencing (Eurofins Genomics). .. Plasmids with the correct sequence were used to transform Rosetta (DE3)pLysS Competent E. coli Cells (Novagen) for recombinant protein production.

    Polymerase Chain Reaction:

    Article Title: Acclimation of Photosynthesis to the Environment 1 acts as a copper-binding superoxide detoxification enzyme.
    Article Snippet: Briefly, pLIC03-APE1 was amplified via PCR using NEB Q5 High-Fidelity 2X Master Mix in 25 μl reaction volumes containing 10 ng DNA template and 0.5 μM of specific primers pairs carrying the new codon (C128S mutation: TGC-AGC; H163A: CAC-GCG; C247S: TGC-AGC); the PCR protocol was set according to the manufacturer's instructions (amplification was done for 25 cycles with 180 s elongation time each) and annealing temperatures were calculated with the online NEB Tm calculator tool. .. After amplification, to remove the initial DNA template (i.e., without the added mutation) and ligate the PCR products, 1 μl of PCR product was incubated at RT for 10 min with NEB 10X KLD (Kinase, Ligase, DpnI) Enzyme Mix in a 10 μl reaction volume before being used to transform NEB 5-alpha competent E. coli cells, for plasmid isolation and Sanger sequencing (Eurofins Genomics). .. Plasmids with the correct sequence were used to transform Rosetta (DE3)pLysS Competent E. coli Cells (Novagen) for recombinant protein production.

    Incubation:

    Article Title: Acclimation of Photosynthesis to the Environment 1 acts as a copper-binding superoxide detoxification enzyme.
    Article Snippet: Briefly, pLIC03-APE1 was amplified via PCR using NEB Q5 High-Fidelity 2X Master Mix in 25 μl reaction volumes containing 10 ng DNA template and 0.5 μM of specific primers pairs carrying the new codon (C128S mutation: TGC-AGC; H163A: CAC-GCG; C247S: TGC-AGC); the PCR protocol was set according to the manufacturer's instructions (amplification was done for 25 cycles with 180 s elongation time each) and annealing temperatures were calculated with the online NEB Tm calculator tool. .. After amplification, to remove the initial DNA template (i.e., without the added mutation) and ligate the PCR products, 1 μl of PCR product was incubated at RT for 10 min with NEB 10X KLD (Kinase, Ligase, DpnI) Enzyme Mix in a 10 μl reaction volume before being used to transform NEB 5-alpha competent E. coli cells, for plasmid isolation and Sanger sequencing (Eurofins Genomics). .. Plasmids with the correct sequence were used to transform Rosetta (DE3)pLysS Competent E. coli Cells (Novagen) for recombinant protein production.

    Plasmid Preparation:

    Article Title: Acclimation of Photosynthesis to the Environment 1 acts as a copper-binding superoxide detoxification enzyme.
    Article Snippet: Briefly, pLIC03-APE1 was amplified via PCR using NEB Q5 High-Fidelity 2X Master Mix in 25 μl reaction volumes containing 10 ng DNA template and 0.5 μM of specific primers pairs carrying the new codon (C128S mutation: TGC-AGC; H163A: CAC-GCG; C247S: TGC-AGC); the PCR protocol was set according to the manufacturer's instructions (amplification was done for 25 cycles with 180 s elongation time each) and annealing temperatures were calculated with the online NEB Tm calculator tool. .. After amplification, to remove the initial DNA template (i.e., without the added mutation) and ligate the PCR products, 1 μl of PCR product was incubated at RT for 10 min with NEB 10X KLD (Kinase, Ligase, DpnI) Enzyme Mix in a 10 μl reaction volume before being used to transform NEB 5-alpha competent E. coli cells, for plasmid isolation and Sanger sequencing (Eurofins Genomics). .. Plasmids with the correct sequence were used to transform Rosetta (DE3)pLysS Competent E. coli Cells (Novagen) for recombinant protein production.

    Isolation:

    Article Title: Acclimation of Photosynthesis to the Environment 1 acts as a copper-binding superoxide detoxification enzyme.
    Article Snippet: Briefly, pLIC03-APE1 was amplified via PCR using NEB Q5 High-Fidelity 2X Master Mix in 25 μl reaction volumes containing 10 ng DNA template and 0.5 μM of specific primers pairs carrying the new codon (C128S mutation: TGC-AGC; H163A: CAC-GCG; C247S: TGC-AGC); the PCR protocol was set according to the manufacturer's instructions (amplification was done for 25 cycles with 180 s elongation time each) and annealing temperatures were calculated with the online NEB Tm calculator tool. .. After amplification, to remove the initial DNA template (i.e., without the added mutation) and ligate the PCR products, 1 μl of PCR product was incubated at RT for 10 min with NEB 10X KLD (Kinase, Ligase, DpnI) Enzyme Mix in a 10 μl reaction volume before being used to transform NEB 5-alpha competent E. coli cells, for plasmid isolation and Sanger sequencing (Eurofins Genomics). .. Plasmids with the correct sequence were used to transform Rosetta (DE3)pLysS Competent E. coli Cells (Novagen) for recombinant protein production.

    DNA Sequencing:

    Article Title: Acclimation of Photosynthesis to the Environment 1 acts as a copper-binding superoxide detoxification enzyme.
    Article Snippet: Briefly, pLIC03-APE1 was amplified via PCR using NEB Q5 High-Fidelity 2X Master Mix in 25 μl reaction volumes containing 10 ng DNA template and 0.5 μM of specific primers pairs carrying the new codon (C128S mutation: TGC-AGC; H163A: CAC-GCG; C247S: TGC-AGC); the PCR protocol was set according to the manufacturer's instructions (amplification was done for 25 cycles with 180 s elongation time each) and annealing temperatures were calculated with the online NEB Tm calculator tool. .. After amplification, to remove the initial DNA template (i.e., without the added mutation) and ligate the PCR products, 1 μl of PCR product was incubated at RT for 10 min with NEB 10X KLD (Kinase, Ligase, DpnI) Enzyme Mix in a 10 μl reaction volume before being used to transform NEB 5-alpha competent E. coli cells, for plasmid isolation and Sanger sequencing (Eurofins Genomics). .. Plasmids with the correct sequence were used to transform Rosetta (DE3)pLysS Competent E. coli Cells (Novagen) for recombinant protein production.



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