gibson assembly kit  (New England Biolabs)


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  • 99
    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, 2020-08
    99/100 stars

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    Related Articles

    Clone Assay:

    Article Title: Molecular analysis of NPAS3 functional domains and variants
    Article Snippet: .. The variant was assembled into digested pDONR221- NPAS3 transcript variant 1 using the Gibson Assembly Cloning Kit (NEB), and recombined into the pcI-HA destination vector. .. HEK 293T cells were purchased from ATCC and cultured in Dulbecco’s Modified Eagle Medium (DMEM) with high glucose (Sigma) in a 37 °C incubator with a humidified 5% CO2 atmosphere.

    Article Title: Aging-like Spontaneous Epigenetic Silencing Facilitates Wnt Activation, Stemness, and BrafV600E-Induced Tumorigenesis
    Article Snippet: .. Puromycin resistance gene in Puro.Cre empty vector (Addgene #17408) was replaced by hygromycin resistance gene PCR amplified from pBABE-hygrohTERT (Addgene #1773) using primer listed below (Hyg F and R) by Gibson Assembly® cloning kit (NEB® #E5510S) generating Lenti-cre vector. .. Subsequently, Cre in the Lenti-cre vector was deleted by Gibson Assembly® cloning kit generating Lenti-empty vector using primers (Cre deletion F and R).

    Article Title: Intra-Seasonal Antibody Repertoire Analysis of a Patient Immunized with an MF59®-adjuvanted Pandemic 2009 H1N1 Vaccine
    Article Snippet: .. Linear DNA cassettes for Ig VH , Vκ , and Vλ segments were synthesized as GBIocks at Integrated DNA Technologies and cloned into pRS5α expressing full-length IgG1 heavy, kappa light, or lambda light chains where appropriate, by Gibson assembly (New England Biolabs, E5510S) according to the recommended protocol. .. Variable regions of Ig chains were sequence verified.

    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: Simple Protocol for Generating and Genotyping Genome-Edited Mice With CRISPR-Cas9 Reagents.
    Article Snippet: .. PCR products can be cloned into pGEM T-Easy Vector (Promega) or pELuc (Lunyak et al., 2007) using the Gibson Assembly® Cloning Kit (New England Biolabs, procedure described in Gibson et al., 2009). .. DNA primers used for the amplification of genomic sequences include a 5′ adapter sequenceFernández et al. 14 of 18 Current Protocols in Mouse Biology Figure 7 Many positive genome-edited founder mice can be detected by RFLP analyses upon digestion with HinfI (left).

    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: A role for 2-Cys peroxiredoxins in facilitating cytosolic protein thiol oxidation
    Article Snippet: .. Open reading frames and the streptavidin binding peptide (SBP) tag were amplified by PCR and ligated into pcDNA3.1(−) using the Gibson Assembly Cloning Kit (New England BioLabs). .. Mutations were introduced by using the QuikChange Site-Directed Mutagenesis Kit (Agilent).

    Amplification:

    Article Title: Aging-like Spontaneous Epigenetic Silencing Facilitates Wnt Activation, Stemness, and BrafV600E-Induced Tumorigenesis
    Article Snippet: .. Puromycin resistance gene in Puro.Cre empty vector (Addgene #17408) was replaced by hygromycin resistance gene PCR amplified from pBABE-hygrohTERT (Addgene #1773) using primer listed below (Hyg F and R) by Gibson Assembly® cloning kit (NEB® #E5510S) generating Lenti-cre vector. .. Subsequently, Cre in the Lenti-cre vector was deleted by Gibson Assembly® cloning kit generating Lenti-empty vector using primers (Cre deletion F and R).

    Article Title: A role for 2-Cys peroxiredoxins in facilitating cytosolic protein thiol oxidation
    Article Snippet: .. Open reading frames and the streptavidin binding peptide (SBP) tag were amplified by PCR and ligated into pcDNA3.1(−) using the Gibson Assembly Cloning Kit (New England BioLabs). .. Mutations were introduced by using the QuikChange Site-Directed Mutagenesis Kit (Agilent).

    Expressing:

    Article Title: Intra-Seasonal Antibody Repertoire Analysis of a Patient Immunized with an MF59®-adjuvanted Pandemic 2009 H1N1 Vaccine
    Article Snippet: .. Linear DNA cassettes for Ig VH , Vκ , and Vλ segments were synthesized as GBIocks at Integrated DNA Technologies and cloned into pRS5α expressing full-length IgG1 heavy, kappa light, or lambda light chains where appropriate, by Gibson assembly (New England Biolabs, E5510S) according to the recommended protocol. .. Variable regions of Ig chains were sequence verified.

    Synthesized:

    Article Title: Intra-Seasonal Antibody Repertoire Analysis of a Patient Immunized with an MF59®-adjuvanted Pandemic 2009 H1N1 Vaccine
    Article Snippet: .. Linear DNA cassettes for Ig VH , Vκ , and Vλ segments were synthesized as GBIocks at Integrated DNA Technologies and cloned into pRS5α expressing full-length IgG1 heavy, kappa light, or lambda light chains where appropriate, by Gibson assembly (New England Biolabs, E5510S) according to the recommended protocol. .. Variable regions of Ig chains were sequence verified.

    Variant Assay:

    Article Title: Molecular analysis of NPAS3 functional domains and variants
    Article Snippet: .. The variant was assembled into digested pDONR221- NPAS3 transcript variant 1 using the Gibson Assembly Cloning Kit (NEB), and recombined into the pcI-HA destination vector. .. HEK 293T cells were purchased from ATCC and cultured in Dulbecco’s Modified Eagle Medium (DMEM) with high glucose (Sigma) in a 37 °C incubator with a humidified 5% CO2 atmosphere.

    Polymerase Chain Reaction:

    Article Title: Aging-like Spontaneous Epigenetic Silencing Facilitates Wnt Activation, Stemness, and BrafV600E-Induced Tumorigenesis
    Article Snippet: .. Puromycin resistance gene in Puro.Cre empty vector (Addgene #17408) was replaced by hygromycin resistance gene PCR amplified from pBABE-hygrohTERT (Addgene #1773) using primer listed below (Hyg F and R) by Gibson Assembly® cloning kit (NEB® #E5510S) generating Lenti-cre vector. .. Subsequently, Cre in the Lenti-cre vector was deleted by Gibson Assembly® cloning kit generating Lenti-empty vector using primers (Cre deletion F and R).

    Article Title: Simple Protocol for Generating and Genotyping Genome-Edited Mice With CRISPR-Cas9 Reagents.
    Article Snippet: .. PCR products can be cloned into pGEM T-Easy Vector (Promega) or pELuc (Lunyak et al., 2007) using the Gibson Assembly® Cloning Kit (New England Biolabs, procedure described in Gibson et al., 2009). .. DNA primers used for the amplification of genomic sequences include a 5′ adapter sequenceFernández et al. 14 of 18 Current Protocols in Mouse Biology Figure 7 Many positive genome-edited founder mice can be detected by RFLP analyses upon digestion with HinfI (left).

    Article Title: A role for 2-Cys peroxiredoxins in facilitating cytosolic protein thiol oxidation
    Article Snippet: .. Open reading frames and the streptavidin binding peptide (SBP) tag were amplified by PCR and ligated into pcDNA3.1(−) using the Gibson Assembly Cloning Kit (New England BioLabs). .. Mutations were introduced by using the QuikChange Site-Directed Mutagenesis Kit (Agilent).

    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). ..

    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). ..

    Binding Assay:

    Article Title: A role for 2-Cys peroxiredoxins in facilitating cytosolic protein thiol oxidation
    Article Snippet: .. Open reading frames and the streptavidin binding peptide (SBP) tag were amplified by PCR and ligated into pcDNA3.1(−) using the Gibson Assembly Cloning Kit (New England BioLabs). .. Mutations were introduced by using the QuikChange Site-Directed Mutagenesis Kit (Agilent).

    Plasmid Preparation:

    Article Title: Molecular analysis of NPAS3 functional domains and variants
    Article Snippet: .. The variant was assembled into digested pDONR221- NPAS3 transcript variant 1 using the Gibson Assembly Cloning Kit (NEB), and recombined into the pcI-HA destination vector. .. HEK 293T cells were purchased from ATCC and cultured in Dulbecco’s Modified Eagle Medium (DMEM) with high glucose (Sigma) in a 37 °C incubator with a humidified 5% CO2 atmosphere.

    Article Title: Aging-like Spontaneous Epigenetic Silencing Facilitates Wnt Activation, Stemness, and BrafV600E-Induced Tumorigenesis
    Article Snippet: .. Puromycin resistance gene in Puro.Cre empty vector (Addgene #17408) was replaced by hygromycin resistance gene PCR amplified from pBABE-hygrohTERT (Addgene #1773) using primer listed below (Hyg F and R) by Gibson Assembly® cloning kit (NEB® #E5510S) generating Lenti-cre vector. .. Subsequently, Cre in the Lenti-cre vector was deleted by Gibson Assembly® cloning kit generating Lenti-empty vector using primers (Cre deletion F and R).

    Article Title: Simple Protocol for Generating and Genotyping Genome-Edited Mice With CRISPR-Cas9 Reagents.
    Article Snippet: .. PCR products can be cloned into pGEM T-Easy Vector (Promega) or pELuc (Lunyak et al., 2007) using the Gibson Assembly® Cloning Kit (New England Biolabs, procedure described in Gibson et al., 2009). .. DNA primers used for the amplification of genomic sequences include a 5′ adapter sequenceFernández et al. 14 of 18 Current Protocols in Mouse Biology Figure 7 Many positive genome-edited founder mice can be detected by RFLP analyses upon digestion with HinfI (left).

    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, 2020-08
    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, 2020-08
    99/100 stars
      Buy from Supplier

    99
    New England Biolabs hifi dna assembly cloning 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.
    Hifi Dna Assembly Cloning Kit, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 68 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hifi dna assembly cloning kit/product/New England Biolabs
    Average 99 stars, based on 68 article reviews
    Price from $9.99 to $1999.99
    hifi dna assembly cloning kit - by Bioz Stars, 2020-08
    99/100 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