kld  (New England Biolabs)


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  • 99
    Name:
    KLD Enzyme Mix
    Description:
    KLD Enzyme Mix 25 rxns
    Catalog Number:
    M0554S
    Price:
    312
    Category:
    PCR Mutagenesis Kits
    Size:
    25 rxns
    Buy from Supplier


    Structured Review

    New England Biolabs kld
    KLD Enzyme Mix
    KLD Enzyme Mix 25 rxns
    https://www.bioz.com/result/kld/product/New England Biolabs
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    kld - by Bioz Stars, 2021-06
    99/100 stars

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    Related Articles

    Sequencing:

    Article Title: An engineered ScCas9 with broad PAM range and high specificity and activity.
    Article Snippet: CRISPR enzymes require a protospacer-adjacent motif (PAM) near the target cleavage site, constraining the sequences accessible for editing. .. CRISPR enzymes require a protospacer-adjacent motif (PAM) near the target cleavage site, constraining the sequences accessible for editing. .. CRISPR enzymes require a protospacer-adjacent motif (PAM) near the target cleavage site, constraining the sequences accessible for editing.

    Polymerase Chain Reaction:

    Article Title: An engineered ScCas9 with broad PAM range and high specificity and activity.
    Article Snippet: CRISPR enzymes require a protospacer-adjacent motif (PAM) near the target cleavage site, constraining the sequences accessible for editing. .. CRISPR enzymes require a protospacer-adjacent motif (PAM) near the target cleavage site, constraining the sequences accessible for editing. .. CRISPR enzymes require a protospacer-adjacent motif (PAM) near the target cleavage site, constraining the sequences accessible for editing.

    Article Title: A Single Salt Bridge in VIM-20 Increases Protein Stability and Antibiotic Resistance under Low-Zinc Conditions
    Article Snippet: An expression plasmid for VIM-20, which harbors the H229R mutation, was generated by site-directed mutagenesis using a forward primer (5′-TATTCAGCAACGTTACCCGGAAGCGCAATTC-3′), a reverse primer (5′-CGTTCGATGCTGGTCGGC-3′), and the pET-28a(+)-TEV-VIM-2 plasmid as the template. .. After the PCR, the amplified sequences were added directly to a kinase-ligase-DpnI enzyme mix (NEB) for rapid, room-temperature circularization and template removal. .. A 2-μl aliquot of the ligation mixture was used to transform 30 μl of E. coli DH5α chemically competent cells (Lucigen), and the transformation mixture was spread onto a lysogeny broth (LB) plate containing 50 μg/ml kanamycin.

    Article Title: Post-Translational Sortase-Mediated Attachment of High-Strength Force Spectroscopy Handles
    Article Snippet: Cloning Modified pET28a plasmids encoding for ybbR-His-XylanaseT6(T129C) (Geobacillus stearothermophilus )-Doc3 (R. flavefaciens ), ybbR-His-sfGFP-DocI (Clostridium thermocellum ), and Titin-Ig domains (repeats 27 to 32, repeat 34, human) were used as templates for polymerase chain reaction (PCR) with subsequent reconstitution by Gibson assembly. .. The previously reported d59 sortase(P94R/D160N/D165A/K190E/K196T) mutant was created by introducing the mutations via overlap extension PCR followed by ligating the linearized plasmid using Kinase–Ligase–DpnI (KLD) enzyme mix and KLD reaction buffer from the Q5 site-directed mutagenesis kit (New England Biolabs, MA, USA). .. The chemically competent E. coli DH5-α cells were transformed [Life Technologies GmbH, Frankfurt, Germany; 30 min on ice, 30 s heat shock at 42 °C followed by 37 °C for 1 h in a super optimal broth with catabolite repression medium] and plated on kanamycin-supplemented agar plates.

    Article Title: Resolving dual binding conformations of cellulosome cohesin-dockerin complexes using single-molecule force spectroscopy
    Article Snippet: For insertion of the (GS)4 and (GS)8 linkers into the Doc domain, exponential amplification with primers bearing coding sequences for the inserts at their 5’-ends was performed with a Phusion High-Fidelity DNA polymerase (New England Biolabs, MA). .. PCR products were then blunt end ligated using KLD Enzyme Mix and KLD Reaction Buffer from the Q5 site directed mutagenesis kit (New England Biolabs, MA). .. The modified DNA constructs were used to transform Escherichia coli DH5-alpha cells, grown on kanamycin-containing agar plates and subsequently screened.

    Article Title: The cryo-EM structure of the bacterial flagellum cap complex suggests a molecular mechanism for filament elongation
    Article Snippet: Briefly, the fliD complementation plasmid was amplified by PCR with divergent primers containing targeted nucleotide substitutions in the forward primer (listed in Supplementary Table ). .. An aliquot of the linear PCR product was treated with the KLD enzyme mix to circularize the mutated plasmid while degrading any residual template. .. The treated plasmids were transformed into E. coli DH5ɑ and transformants selected by chloramphenicol resistance.

    Amplification:

    Article Title: An engineered ScCas9 with broad PAM range and high specificity and activity.
    Article Snippet: CRISPR enzymes require a protospacer-adjacent motif (PAM) near the target cleavage site, constraining the sequences accessible for editing. .. CRISPR enzymes require a protospacer-adjacent motif (PAM) near the target cleavage site, constraining the sequences accessible for editing. .. CRISPR enzymes require a protospacer-adjacent motif (PAM) near the target cleavage site, constraining the sequences accessible for editing.

    Article Title: A Single Salt Bridge in VIM-20 Increases Protein Stability and Antibiotic Resistance under Low-Zinc Conditions
    Article Snippet: An expression plasmid for VIM-20, which harbors the H229R mutation, was generated by site-directed mutagenesis using a forward primer (5′-TATTCAGCAACGTTACCCGGAAGCGCAATTC-3′), a reverse primer (5′-CGTTCGATGCTGGTCGGC-3′), and the pET-28a(+)-TEV-VIM-2 plasmid as the template. .. After the PCR, the amplified sequences were added directly to a kinase-ligase-DpnI enzyme mix (NEB) for rapid, room-temperature circularization and template removal. .. A 2-μl aliquot of the ligation mixture was used to transform 30 μl of E. coli DH5α chemically competent cells (Lucigen), and the transformation mixture was spread onto a lysogeny broth (LB) plate containing 50 μg/ml kanamycin.

    Construct:

    Article Title: Development of a Formaldehyde Biosensor with Application to Synthetic Methylotrophy
    Article Snippet: Pathway optimization assays were conducted in E. coli MG1655(DE3) ( ). pBbS2k-RFP was a gift from Jay Keasling (Addgene plasmid #35330). pETM6-mCherry was a gift from Mattheos Koffas (Addgene plasmid #66534). .. Plasmids were constructed using USER cloning, Gibson Assembly Master Mix, or KLD Enzyme Mix (NEB). .. DNA fragments were generated by PCR with the primers listed in .

    Clone Assay:

    Article Title: Development of a Formaldehyde Biosensor with Application to Synthetic Methylotrophy
    Article Snippet: Pathway optimization assays were conducted in E. coli MG1655(DE3) ( ). pBbS2k-RFP was a gift from Jay Keasling (Addgene plasmid #35330). pETM6-mCherry was a gift from Mattheos Koffas (Addgene plasmid #66534). .. Plasmids were constructed using USER cloning, Gibson Assembly Master Mix, or KLD Enzyme Mix (NEB). .. DNA fragments were generated by PCR with the primers listed in .

    Mutagenesis:

    Article Title: Post-Translational Sortase-Mediated Attachment of High-Strength Force Spectroscopy Handles
    Article Snippet: Cloning Modified pET28a plasmids encoding for ybbR-His-XylanaseT6(T129C) (Geobacillus stearothermophilus )-Doc3 (R. flavefaciens ), ybbR-His-sfGFP-DocI (Clostridium thermocellum ), and Titin-Ig domains (repeats 27 to 32, repeat 34, human) were used as templates for polymerase chain reaction (PCR) with subsequent reconstitution by Gibson assembly. .. The previously reported d59 sortase(P94R/D160N/D165A/K190E/K196T) mutant was created by introducing the mutations via overlap extension PCR followed by ligating the linearized plasmid using Kinase–Ligase–DpnI (KLD) enzyme mix and KLD reaction buffer from the Q5 site-directed mutagenesis kit (New England Biolabs, MA, USA). .. The chemically competent E. coli DH5-α cells were transformed [Life Technologies GmbH, Frankfurt, Germany; 30 min on ice, 30 s heat shock at 42 °C followed by 37 °C for 1 h in a super optimal broth with catabolite repression medium] and plated on kanamycin-supplemented agar plates.

    Article Title: Resolving dual binding conformations of cellulosome cohesin-dockerin complexes using single-molecule force spectroscopy
    Article Snippet: For insertion of the (GS)4 and (GS)8 linkers into the Doc domain, exponential amplification with primers bearing coding sequences for the inserts at their 5’-ends was performed with a Phusion High-Fidelity DNA polymerase (New England Biolabs, MA). .. PCR products were then blunt end ligated using KLD Enzyme Mix and KLD Reaction Buffer from the Q5 site directed mutagenesis kit (New England Biolabs, MA). .. The modified DNA constructs were used to transform Escherichia coli DH5-alpha cells, grown on kanamycin-containing agar plates and subsequently screened.

    Plasmid Preparation:

    Article Title: Post-Translational Sortase-Mediated Attachment of High-Strength Force Spectroscopy Handles
    Article Snippet: Cloning Modified pET28a plasmids encoding for ybbR-His-XylanaseT6(T129C) (Geobacillus stearothermophilus )-Doc3 (R. flavefaciens ), ybbR-His-sfGFP-DocI (Clostridium thermocellum ), and Titin-Ig domains (repeats 27 to 32, repeat 34, human) were used as templates for polymerase chain reaction (PCR) with subsequent reconstitution by Gibson assembly. .. The previously reported d59 sortase(P94R/D160N/D165A/K190E/K196T) mutant was created by introducing the mutations via overlap extension PCR followed by ligating the linearized plasmid using Kinase–Ligase–DpnI (KLD) enzyme mix and KLD reaction buffer from the Q5 site-directed mutagenesis kit (New England Biolabs, MA, USA). .. The chemically competent E. coli DH5-α cells were transformed [Life Technologies GmbH, Frankfurt, Germany; 30 min on ice, 30 s heat shock at 42 °C followed by 37 °C for 1 h in a super optimal broth with catabolite repression medium] and plated on kanamycin-supplemented agar plates.

    Article Title: The cryo-EM structure of the bacterial flagellum cap complex suggests a molecular mechanism for filament elongation
    Article Snippet: Briefly, the fliD complementation plasmid was amplified by PCR with divergent primers containing targeted nucleotide substitutions in the forward primer (listed in Supplementary Table ). .. An aliquot of the linear PCR product was treated with the KLD enzyme mix to circularize the mutated plasmid while degrading any residual template. .. The treated plasmids were transformed into E. coli DH5ɑ and transformants selected by chloramphenicol resistance.

    Transformation Assay:

    Article Title: Clamping, bending, and twisting inter-domain motions in the misfold-recognizing portion of UDP-glucose: Glycoprotein glucosyltransferase
    Article Snippet: Kinase, Ligase & DpnI (KLD) treatment: 1 μL of PCR product was mixed with 5 μL of 2X KLD Reaction buffer, 1 μL of 10X KLD Enzyme Mix (both from NEB) and 3 μL of nuclease-free water. .. The KLD reaction mixture was used to transform E . coli DH-5α chemically competent cells (NEB) using the following protocol: 5 μL of KLD reaction mix were added to a tube of thawed DH-5α competent E . coli cells on ice, and mixed gently for a few seconds; after transformation, the bacteria were incubated on ice for 30 minutes, heat shocked at 42 °C for 30 seconds and incubated on ice again for 5 minutes. ..

    Incubation:

    Article Title: Clamping, bending, and twisting inter-domain motions in the misfold-recognizing portion of UDP-glucose: Glycoprotein glucosyltransferase
    Article Snippet: Kinase, Ligase & DpnI (KLD) treatment: 1 μL of PCR product was mixed with 5 μL of 2X KLD Reaction buffer, 1 μL of 10X KLD Enzyme Mix (both from NEB) and 3 μL of nuclease-free water. .. The KLD reaction mixture was used to transform E . coli DH-5α chemically competent cells (NEB) using the following protocol: 5 μL of KLD reaction mix were added to a tube of thawed DH-5α competent E . coli cells on ice, and mixed gently for a few seconds; after transformation, the bacteria were incubated on ice for 30 minutes, heat shocked at 42 °C for 30 seconds and incubated on ice again for 5 minutes. ..

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  • 99
    New England Biolabs kld reaction mixture
    Kld Reaction Mixture, 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/kld reaction mixture/product/New England Biolabs
    Average 99 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    kld reaction mixture - by Bioz Stars, 2021-06
    99/100 stars
      Buy from Supplier

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