gibson assembly kit  (New England Biolabs)


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    Name:
    Gibson Assembly Cloning Kit
    Description:
    Gibson Assembly Cloning Kit 10 rxns
    Catalog Number:
    e5510s
    Price:
    195
    Size:
    10 rxns
    Category:
    Cloning and Expression Systems
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    Structured Review

    New England Biolabs gibson assembly kit
    Gibson Assembly Cloning Kit
    Gibson Assembly Cloning Kit 10 rxns
    https://www.bioz.com/result/gibson assembly kit/product/New England Biolabs
    Average 99 stars, based on 189 article reviews
    Price from $9.99 to $1999.99
    gibson assembly kit - by Bioz Stars, 2020-11
    99/100 stars

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    Related Articles

    Clone Assay:

    Article Title: Enhancement of Heat Stability and Kinetic Parameters of the Maize Endosperm ADP-Glucose Pyrophosphorylase by Mutagenesis of Amino Acids in the Small Subunit With High B Factors
    Article Snippet: .. Mutants were prepared using the Gibson Assembly Cloning Kit (NEB) to combine PCR products with the vector. .. PCR products for single site mutants were prepared using pMONc Bt2 as template.

    Article Title: Increased levels of RECQ5 shift DNA repair from canonical to alternative pathways
    Article Snippet: .. Briefly, 60-mer oligos were annealed and extended then cloned using the Gibson Assembly® Cloning Kit (NEB). .. The sgRNAs used to target the TL reporter were (protospacer-associated motifs capitalized): g1: 5′-gtgtccggcctcgaccgtgAGG g2: 5′-CCGtgaggaggtttctctgtaa g9: 5′-aaagctaagagctcacctaCGG The dsDNA donor plasmid for correction of the TL reporter, pCVL SFFV d14GFP, was previously described ( ).

    Article Title: Laboratory evolution for forced glucose-xylose co-consumption enables identification of mutations that improve mixed-sugar fermentation by xylose-fermenting Saccharomyces cerevisiae
    Article Snippet: .. Plasmid assembly was performed in vitro with a Gibson Assembly Cloning kit (New England Biolabs, Ipswich, MA), following the supplier's guidelines, or in vivo by transformation of plasmid fragments into yeast cells (Kuijpers et al. ). .. For all constructs, correct assembly was confirmed by diagnostic PCR with DreamTaq polymerase (Thermo-Scientific), following the manufacturer's protocol.

    Article Title: Mild hydrostatic pressure triggers oxidative responses in Escherichia coli
    Article Snippet: .. Gibson assembly reactions were performed using NEB Gibson Assembly® Cloning Kit (NEB, UK) according to the manufacturer’s instructions. .. The reaction mixture then was transformed into competent cells o f E . coli NEB 5α, and selected on LB agar plate containing 12.5 μg/ml chloramphenicol.

    Article Title: Transcription Interference and ORF Nature Strongly Affect Promoter Strength in a Reconstituted Metabolic Pathway
    Article Snippet: .. The library of promoter-ORF-terminator associations was subsequently assembled by Gibson cloning method (Gibson et al., ) in a Sac I/Xho I linearized pRS426 (URA3 selection marker) (Gibson Assembly Cloning Kit, NEB, Ipswich, MA, USA). ..

    Article Title: Optimizing anaerobic growth rate and fermentation kinetics in Saccharomyces cerevisiae strains expressing Calvin-cycle enzymes for improved ethanol yield
    Article Snippet: .. The assembly of plasmids pUDR119 and pUDR164 was performed in vitro using the Gibson Assembly Cloning kit (New England Biolabs, Ipswich, MA) following the supplier’s guidelines. .. The assembly was enabled by homologous sequences present at the 5′ and 3′ ends of the PCR-amplified plasmid backbones and inserts.

    In Vivo:

    Article Title: Laboratory evolution for forced glucose-xylose co-consumption enables identification of mutations that improve mixed-sugar fermentation by xylose-fermenting Saccharomyces cerevisiae
    Article Snippet: .. Plasmid assembly was performed in vitro with a Gibson Assembly Cloning kit (New England Biolabs, Ipswich, MA), following the supplier's guidelines, or in vivo by transformation of plasmid fragments into yeast cells (Kuijpers et al. ). .. For all constructs, correct assembly was confirmed by diagnostic PCR with DreamTaq polymerase (Thermo-Scientific), following the manufacturer's protocol.

    Selection:

    Article Title: Transcription Interference and ORF Nature Strongly Affect Promoter Strength in a Reconstituted Metabolic Pathway
    Article Snippet: .. The library of promoter-ORF-terminator associations was subsequently assembled by Gibson cloning method (Gibson et al., ) in a Sac I/Xho I linearized pRS426 (URA3 selection marker) (Gibson Assembly Cloning Kit, NEB, Ipswich, MA, USA). ..

    In Vitro:

    Article Title: Laboratory evolution for forced glucose-xylose co-consumption enables identification of mutations that improve mixed-sugar fermentation by xylose-fermenting Saccharomyces cerevisiae
    Article Snippet: .. Plasmid assembly was performed in vitro with a Gibson Assembly Cloning kit (New England Biolabs, Ipswich, MA), following the supplier's guidelines, or in vivo by transformation of plasmid fragments into yeast cells (Kuijpers et al. ). .. For all constructs, correct assembly was confirmed by diagnostic PCR with DreamTaq polymerase (Thermo-Scientific), following the manufacturer's protocol.

    Article Title: Optimizing anaerobic growth rate and fermentation kinetics in Saccharomyces cerevisiae strains expressing Calvin-cycle enzymes for improved ethanol yield
    Article Snippet: .. The assembly of plasmids pUDR119 and pUDR164 was performed in vitro using the Gibson Assembly Cloning kit (New England Biolabs, Ipswich, MA) following the supplier’s guidelines. .. The assembly was enabled by homologous sequences present at the 5′ and 3′ ends of the PCR-amplified plasmid backbones and inserts.

    Purification:

    Article Title: OSCA/TMEM63 are an evolutionarily conserved family of mechanically activated ion channels
    Article Snippet: .. Then the purified PCR product ligated into the digested plasmid using Gibson assembly kit (NEB). .. The protein sequence downloaded from TAIR matched the sequence obtained from the plant.

    Polymerase Chain Reaction:

    Article Title: Enhancement of Heat Stability and Kinetic Parameters of the Maize Endosperm ADP-Glucose Pyrophosphorylase by Mutagenesis of Amino Acids in the Small Subunit With High B Factors
    Article Snippet: .. Mutants were prepared using the Gibson Assembly Cloning Kit (NEB) to combine PCR products with the vector. .. PCR products for single site mutants were prepared using pMONc Bt2 as template.

    Article Title: OSCA/TMEM63 are an evolutionarily conserved family of mechanically activated ion channels
    Article Snippet: .. Then the purified PCR product ligated into the digested plasmid using Gibson assembly kit (NEB). .. The protein sequence downloaded from TAIR matched the sequence obtained from the plant.

    BAC Assay:

    Article Title: An integrated platform for genome engineering and gene expression perturbation in Plasmodium falciparum
    Article Snippet: .. Materials pJAZZ-OC Not I Vector (Lucigen, Catalog # 43024) BigEasy-TSA Electrocompetent Cells (Lucigen, Catalog # 60224) BAC-Optimized Replicator™ v2.0 Electrocompetent cells (Lucigen, Catalog # 60210) 2x Gibson Assembly Master Mix (NEB, Catalog #E5510) DNA Polymerase I, Large (Klenow) Fragment (New England Biolabs, Catalog# M0210) Restriction enzymes were purchased from New England Biolabs, unless otherwise stated ..

    Marker:

    Article Title: Transcription Interference and ORF Nature Strongly Affect Promoter Strength in a Reconstituted Metabolic Pathway
    Article Snippet: .. The library of promoter-ORF-terminator associations was subsequently assembled by Gibson cloning method (Gibson et al., ) in a Sac I/Xho I linearized pRS426 (URA3 selection marker) (Gibson Assembly Cloning Kit, NEB, Ipswich, MA, USA). ..

    Transformation Assay:

    Article Title: Laboratory evolution for forced glucose-xylose co-consumption enables identification of mutations that improve mixed-sugar fermentation by xylose-fermenting Saccharomyces cerevisiae
    Article Snippet: .. Plasmid assembly was performed in vitro with a Gibson Assembly Cloning kit (New England Biolabs, Ipswich, MA), following the supplier's guidelines, or in vivo by transformation of plasmid fragments into yeast cells (Kuijpers et al. ). .. For all constructs, correct assembly was confirmed by diagnostic PCR with DreamTaq polymerase (Thermo-Scientific), following the manufacturer's protocol.

    Plasmid Preparation:

    Article Title: Enhancement of Heat Stability and Kinetic Parameters of the Maize Endosperm ADP-Glucose Pyrophosphorylase by Mutagenesis of Amino Acids in the Small Subunit With High B Factors
    Article Snippet: .. Mutants were prepared using the Gibson Assembly Cloning Kit (NEB) to combine PCR products with the vector. .. PCR products for single site mutants were prepared using pMONc Bt2 as template.

    Article Title: Laboratory evolution for forced glucose-xylose co-consumption enables identification of mutations that improve mixed-sugar fermentation by xylose-fermenting Saccharomyces cerevisiae
    Article Snippet: .. Plasmid assembly was performed in vitro with a Gibson Assembly Cloning kit (New England Biolabs, Ipswich, MA), following the supplier's guidelines, or in vivo by transformation of plasmid fragments into yeast cells (Kuijpers et al. ). .. For all constructs, correct assembly was confirmed by diagnostic PCR with DreamTaq polymerase (Thermo-Scientific), following the manufacturer's protocol.

    Article Title: OSCA/TMEM63 are an evolutionarily conserved family of mechanically activated ion channels
    Article Snippet: .. Then the purified PCR product ligated into the digested plasmid using Gibson assembly kit (NEB). .. The protein sequence downloaded from TAIR matched the sequence obtained from the plant.

    Article Title: An integrated platform for genome engineering and gene expression perturbation in Plasmodium falciparum
    Article Snippet: .. Materials pJAZZ-OC Not I Vector (Lucigen, Catalog # 43024) BigEasy-TSA Electrocompetent Cells (Lucigen, Catalog # 60224) BAC-Optimized Replicator™ v2.0 Electrocompetent cells (Lucigen, Catalog # 60210) 2x Gibson Assembly Master Mix (NEB, Catalog #E5510) DNA Polymerase I, Large (Klenow) Fragment (New England Biolabs, Catalog# M0210) Restriction enzymes were purchased from New England Biolabs, unless otherwise stated ..

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  • 99
    New England Biolabs gibson assembly cloning kit
    Proof-of-concept to establish successful transfer of large DNA fragments containing interspersed regions of AT-rich regulatory elements to a linear vector framework. (A) . The schematic shows 7.5 kb and 9.5 kb fragments to be released from extant circular vectors pSN372 and pMG1847, respectively, for <t>assembly</t> into linear vectors. These fragments contain TetR- or TetR-DOZI-based translation regulation modules and a transcriptional unit in which expression of a FLuc reporter CDS is translationally controlled by TetR aptamers located in either the 5’- UTR only or both 5’- and 3’- UTR s. (B) Strategy used to transfer the respective pSN372- and pMG1847-derived fragments into linear plasmids. The original pJAZZ-OC vector (Lucigen) was modified with a <t>multi-cloning</t> site gene block to create pSwing. To facilitate <t>Gibson</t> assembly, pSwing can be digested with restriction enzymes to expose regions homologous to cut pSN372- and pMG1847-derived fragments (red and green). (C) . Restriction digestion analysis confirming proper topological assembly of pSwing, pSN372L and pSN1847L. For pSN1847L, several plasmids that do not contain the expected insert, and likely corresponding to pSwing, are indicated in red font.
    Gibson Assembly Cloning Kit, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 217 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/gibson assembly cloning kit/product/New England Biolabs
    Average 99 stars, based on 217 article reviews
    Price from $9.99 to $1999.99
    gibson assembly cloning kit - by Bioz Stars, 2020-11
    99/100 stars
      Buy from Supplier

    99
    New England Biolabs hifi dna assembly
    Agarose gel electrophoresis of the three assembled genes (a) The first lane contains the GFP gene assembled from 27 dsDNA fragments showing up at the expected 757 bp length. The second lane contains the kanamycin resistance gene assembled from 28 dsDNA fragments showing up at the expected 953 bp length. The third lane contains the tetracycline resistance gene assembled from 45 dsDNA fragments at the expected 1254 bp length. All assemblies were performed using Gibson Assembly master mix. (b) The same assemblies performed using the NEBuilder <t>HiFi</t> <t>DNA</t> Assembly master mix. The agarose gel is stained with ethidium bromide.
    Hifi Dna Assembly, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 57 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hifi dna assembly/product/New England Biolabs
    Average 99 stars, based on 57 article reviews
    Price from $9.99 to $1999.99
    hifi dna assembly - by Bioz Stars, 2020-11
    99/100 stars
      Buy from Supplier

    94
    New England Biolabs gibson assembly kit
    Agarose gel electrophoresis of the three assembled genes (a) The first lane contains the GFP gene assembled from 27 dsDNA fragments showing up at the expected 757 bp length. The second lane contains the kanamycin resistance gene assembled from 28 dsDNA fragments showing up at the expected 953 bp length. The third lane contains the tetracycline resistance gene assembled from 45 dsDNA fragments at the expected 1254 bp length. All assemblies were performed using Gibson Assembly master mix. (b) The same assemblies performed using the NEBuilder <t>HiFi</t> <t>DNA</t> Assembly master mix. The agarose gel is stained with ethidium bromide.
    Gibson Assembly Kit, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 94/100, based on 189 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/gibson assembly kit/product/New England Biolabs
    Average 94 stars, based on 189 article reviews
    Price from $9.99 to $1999.99
    gibson assembly kit - by Bioz Stars, 2020-11
    94/100 stars
      Buy from Supplier

    Image Search Results


    Proof-of-concept to establish successful transfer of large DNA fragments containing interspersed regions of AT-rich regulatory elements to a linear vector framework. (A) . The schematic shows 7.5 kb and 9.5 kb fragments to be released from extant circular vectors pSN372 and pMG1847, respectively, for assembly into linear vectors. These fragments contain TetR- or TetR-DOZI-based translation regulation modules and a transcriptional unit in which expression of a FLuc reporter CDS is translationally controlled by TetR aptamers located in either the 5’- UTR only or both 5’- and 3’- UTR s. (B) Strategy used to transfer the respective pSN372- and pMG1847-derived fragments into linear plasmids. The original pJAZZ-OC vector (Lucigen) was modified with a multi-cloning site gene block to create pSwing. To facilitate Gibson assembly, pSwing can be digested with restriction enzymes to expose regions homologous to cut pSN372- and pMG1847-derived fragments (red and green). (C) . Restriction digestion analysis confirming proper topological assembly of pSwing, pSN372L and pSN1847L. For pSN1847L, several plasmids that do not contain the expected insert, and likely corresponding to pSwing, are indicated in red font.

    Journal: bioRxiv

    Article Title: An integrated platform for genome engineering and gene expression perturbation in Plasmodium falciparum

    doi: 10.1101/816504

    Figure Lengend Snippet: Proof-of-concept to establish successful transfer of large DNA fragments containing interspersed regions of AT-rich regulatory elements to a linear vector framework. (A) . The schematic shows 7.5 kb and 9.5 kb fragments to be released from extant circular vectors pSN372 and pMG1847, respectively, for assembly into linear vectors. These fragments contain TetR- or TetR-DOZI-based translation regulation modules and a transcriptional unit in which expression of a FLuc reporter CDS is translationally controlled by TetR aptamers located in either the 5’- UTR only or both 5’- and 3’- UTR s. (B) Strategy used to transfer the respective pSN372- and pMG1847-derived fragments into linear plasmids. The original pJAZZ-OC vector (Lucigen) was modified with a multi-cloning site gene block to create pSwing. To facilitate Gibson assembly, pSwing can be digested with restriction enzymes to expose regions homologous to cut pSN372- and pMG1847-derived fragments (red and green). (C) . Restriction digestion analysis confirming proper topological assembly of pSwing, pSN372L and pSN1847L. For pSN1847L, several plasmids that do not contain the expected insert, and likely corresponding to pSwing, are indicated in red font.

    Article Snippet: Materials pJAZZ-OC Not I Vector (Lucigen, Catalog # 43024) BigEasy-TSA Electrocompetent Cells (Lucigen, Catalog # 60224) BAC-Optimized Replicator™ v2.0 Electrocompetent cells (Lucigen, Catalog # 60210) 2x Gibson Assembly Master Mix (NEB, Catalog #E5510) DNA Polymerase I, Large (Klenow) Fragment (New England Biolabs, Catalog# M0210) Restriction enzymes were purchased from New England Biolabs, unless otherwise stated

    Techniques: Plasmid Preparation, Expressing, Derivative Assay, Modification, Clone Assay, Blocking Assay

    Average URA3 mRNA level in four conditions . Analyses were performed on set 2 or set 1 (with or without t HMG1 expression, respectively), in glucose (blue bars) or ethanol (red bars) metabolism. Results were normalized with URA3 signal in strain H for strains from set 1, and with URA3 signal in strain H +H for strains from set 2. Error bars represent 1 SD.

    Journal: Frontiers in Bioengineering and Biotechnology

    Article Title: Transcription Interference and ORF Nature Strongly Affect Promoter Strength in a Reconstituted Metabolic Pathway

    doi: 10.3389/fbioe.2015.00021

    Figure Lengend Snippet: Average URA3 mRNA level in four conditions . Analyses were performed on set 2 or set 1 (with or without t HMG1 expression, respectively), in glucose (blue bars) or ethanol (red bars) metabolism. Results were normalized with URA3 signal in strain H for strains from set 1, and with URA3 signal in strain H +H for strains from set 2. Error bars represent 1 SD.

    Article Snippet: The library of promoter-ORF-terminator associations was subsequently assembled by Gibson cloning method (Gibson et al., ) in a Sac I/Xho I linearized pRS426 (URA3 selection marker) (Gibson Assembly Cloning Kit, NEB, Ipswich, MA, USA).

    Techniques: Expressing

    Agarose gel electrophoresis of the three assembled genes (a) The first lane contains the GFP gene assembled from 27 dsDNA fragments showing up at the expected 757 bp length. The second lane contains the kanamycin resistance gene assembled from 28 dsDNA fragments showing up at the expected 953 bp length. The third lane contains the tetracycline resistance gene assembled from 45 dsDNA fragments at the expected 1254 bp length. All assemblies were performed using Gibson Assembly master mix. (b) The same assemblies performed using the NEBuilder HiFi DNA Assembly master mix. The agarose gel is stained with ethidium bromide.

    Journal: PLoS ONE

    Article Title: Rational Design of High-Number dsDNA Fragments Based on Thermodynamics for the Construction of Full-Length Genes in a Single Reaction

    doi: 10.1371/journal.pone.0145682

    Figure Lengend Snippet: Agarose gel electrophoresis of the three assembled genes (a) The first lane contains the GFP gene assembled from 27 dsDNA fragments showing up at the expected 757 bp length. The second lane contains the kanamycin resistance gene assembled from 28 dsDNA fragments showing up at the expected 953 bp length. The third lane contains the tetracycline resistance gene assembled from 45 dsDNA fragments at the expected 1254 bp length. All assemblies were performed using Gibson Assembly master mix. (b) The same assemblies performed using the NEBuilder HiFi DNA Assembly master mix. The agarose gel is stained with ethidium bromide.

    Article Snippet: We decided to investigate if Gibson Assembly kit and a recently derived kit called the NEBuilder HiFi DNA Assembly (NEB #E2621) have any inherent limitation on the number of dsDNA fragments that can be assembled at once.

    Techniques: Agarose Gel Electrophoresis, Staining