gibson assembly master mix  (New England Biolabs)


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  • 99
    Name:
    Gibson Assembly Master Mix
    Description:
    Gibson Assembly Master Mix 50 rxns
    Catalog Number:
    e2611l
    Price:
    649
    Size:
    50 rxns
    Category:
    Cloning and Expression Systems
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    New England Biolabs gibson assembly master mix
    Gibson Assembly Master Mix
    Gibson Assembly Master Mix 50 rxns
    https://www.bioz.com/result/gibson assembly master mix/product/New England Biolabs
    Average 99 stars, based on 452 article reviews
    Price from $9.99 to $1999.99
    gibson assembly master mix - by Bioz Stars, 2020-01
    99/100 stars

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    Related Articles

    Clone Assay:

    Article Title: Reversibly immortalized human umbilical cord-derived mesenchymal stem cells (UC-MSCs) are responsive to BMP9-induced osteogenic and adipogenic differentiation
    Article Snippet: .. All restriction enzymes used for cloning, the Phusion High-Fidelity PCR kit and the Gibson Assembly Master Mix were from New England Biolabs (Ipswich, MA, USA). ..

    Article Title: Chromobodies to Quantify Changes of Endogenous Protein Concentration in Living Cells
    Article Snippet: The expression construct coding for Ubiquitin-Methionine-BC1-TagGFP2 (hereafter referred as Ub-M-BC1-TagGFP2, B ) was generated by Gibson assembly cloning ( ) of the following four fragments: fragment1 - pEGFP-N1 vector backbone digested with NheI and XbaI, fragment 2 - ubiquitin amplified from Ub-M-eGFP, a gift from Nico Dantuma (plasmid # 11938, Addgene, Cambridge, MA) , with the primer set ubiquitin-for and ubiquitin-rev, fragment 3 - BC1-NB amplified from the BC1-CB expression construct described in ( ) with the primer set BC1-for and BC1-rev, fragment 4 - TagGFP2 amplified from the BC1-CB plasmid described in ( ) with the primer set TagGFP2-for and TagGFP2-rev. .. Fragments were assembled using the Gibson-Assembly Master Mix (New England Biolabs GmbH, Frankfurt, Germany) according to the manufacturer's protocol.

    Amplification:

    Article Title: The Essential Role of Hypermutation in Rapid Adaptation to Antibiotic Stress
    Article Snippet: Allelic exchange vectors ( ) were made using the Gibson Assembly master mix (New England BioLabs) with primers listed in . .. Mutant pmrB alleles were amplified from the respective bioreactor-derived endpoint isolate containing the mutation.

    Article Title: Chromobodies to Quantify Changes of Endogenous Protein Concentration in Living Cells
    Article Snippet: The expression construct coding for Ubiquitin-Methionine-BC1-TagGFP2 (hereafter referred as Ub-M-BC1-TagGFP2, B ) was generated by Gibson assembly cloning ( ) of the following four fragments: fragment1 - pEGFP-N1 vector backbone digested with NheI and XbaI, fragment 2 - ubiquitin amplified from Ub-M-eGFP, a gift from Nico Dantuma (plasmid # 11938, Addgene, Cambridge, MA) , with the primer set ubiquitin-for and ubiquitin-rev, fragment 3 - BC1-NB amplified from the BC1-CB expression construct described in ( ) with the primer set BC1-for and BC1-rev, fragment 4 - TagGFP2 amplified from the BC1-CB plasmid described in ( ) with the primer set TagGFP2-for and TagGFP2-rev. .. Fragments were assembled using the Gibson-Assembly Master Mix (New England Biolabs GmbH, Frankfurt, Germany) according to the manufacturer's protocol.

    Article Title: CRISPR Activation Screens Systematically Identify Factors that Drive Neuronal Fate and Reprogramming
    Article Snippet: The Tubb3-IRES-hCD8 vector (pSLQ1760) was assembled with three fragments (5’ homologous arm of Tubb3, IRES-hCD8, and 3’ homologous arm of Tubb3) and a modified pUC19 backbone vector by using Gibson Assembly Master Mix (New England Biolabs). .. Both 5’ and 3’ homology arms were PCR amplified from the genomic DNA extracted from mES cells with Herculase II Fusion DNA polymerase (Agilent).

    Article Title: Chromobodies to Quantify Changes of Endogenous Protein Concentration in Living Cells
    Article Snippet: Fragment 3 - BC1-NB was amplified from the BC1-CB expression construct with the primer set BC1-for and BC1–4GS-rev, fragment 4 - TagGFP2 was amplified from the BC1-TagGFP2 plasmid with the primer set TagGFP2–4GS-for and TagGFP2-rev, fragment 5 - (G4 S)4 -linker was produced by annealing of the two single stranded oligonucleotides 4GS-linker-oligo1 and 4GS-linker-oligo2 with complementary sequences. .. Fragments were assembled using the Gibson-Assembly Master Mix (New England Biolabs).

    Article Title: Engineering proteins for allosteric control by light or ligands
    Article Snippet: Alternatively, Gibson assembly mix (New England Biolabs, cat. no. E2611S) can be used for isothermal assembly of inserted and host plasmid DNA fragments. .. DNA amplification kit (Fischer Scientific, cat. no. FERK0503 and Qiagen, cat. no. 12643) CAUTION Buffers in the kit can cause serious eye and skin irritation.

    Synthesized:

    Article Title: miR-219a-5p Regulates Rorβ During Osteoblast Differentiation and in Age-related Bone Loss
    Article Snippet: .. The entire mouse Rorβ 3’UTR sequence (7279 nucleotide bases in length; Genbank Accession ), was synthesized as four approximately equal-sized gBLOCKs (Integrated DNA Technologies, Coralville, IA, USA) and assembled into the XbaI site of Luc2P-4TO using Gibson Assembly Master Mix (New England BioLabs, Ipswich, MA, USA) to produce Rorβ−3’UTR-Luc. .. To assess the effects of miR-219a-5p on the Rorβ 3’UTR, 100 ng of Rorβ−3’UTR-Luc were cotransfected with 100nM of either negative control or miR-219a-5p miRNA mimics (Qiagen) into primary mouse CalOBs ( n = 6) at approximately 70% confluence using DharmaFECT DUO transfection reagent (Dharmacon), a reagent specifically made for cotransfection of DNA and miRNAs.

    Construct:

    Article Title: miR-219a-5p Regulates Rorβ During Osteoblast Differentiation and in Age-related Bone Loss
    Article Snippet: Paragraph title: Rorβ−3’UTR luciferase report construct and assay ... The entire mouse Rorβ 3’UTR sequence (7279 nucleotide bases in length; Genbank Accession ), was synthesized as four approximately equal-sized gBLOCKs (Integrated DNA Technologies, Coralville, IA, USA) and assembled into the XbaI site of Luc2P-4TO using Gibson Assembly Master Mix (New England BioLabs, Ipswich, MA, USA) to produce Rorβ−3’UTR-Luc.

    Article Title: The Essential Role of Hypermutation in Rapid Adaptation to Antibiotic Stress
    Article Snippet: Point mutants in PAO1 were constructed using the protocol described by Huang and Wilks , with minor modifications. pEX18Gm was used instead of pEX18Tc. .. Allelic exchange vectors ( ) were made using the Gibson Assembly master mix (New England BioLabs) with primers listed in .

    Article Title: Chromobodies to Quantify Changes of Endogenous Protein Concentration in Living Cells
    Article Snippet: Paragraph title: Expression Constructs ... Fragments were assembled using the Gibson-Assembly Master Mix (New England Biolabs GmbH, Frankfurt, Germany) according to the manufacturer's protocol.

    Incubation:

    Article Title: The Essential Role of Hypermutation in Rapid Adaptation to Antibiotic Stress
    Article Snippet: Allelic exchange vectors ( ) were made using the Gibson Assembly master mix (New England BioLabs) with primers listed in . .. Electroporated cells were spread on BHI plus 60 μg/ml gentamicin (Gm) plates and incubated at 37°C for 2 to 3 days.

    Luciferase:

    Article Title: miR-219a-5p Regulates Rorβ During Osteoblast Differentiation and in Age-related Bone Loss
    Article Snippet: Paragraph title: Rorβ−3’UTR luciferase report construct and assay ... The entire mouse Rorβ 3’UTR sequence (7279 nucleotide bases in length; Genbank Accession ), was synthesized as four approximately equal-sized gBLOCKs (Integrated DNA Technologies, Coralville, IA, USA) and assembled into the XbaI site of Luc2P-4TO using Gibson Assembly Master Mix (New England BioLabs, Ipswich, MA, USA) to produce Rorβ−3’UTR-Luc.

    Activity Assay:

    Article Title: miR-219a-5p Regulates Rorβ During Osteoblast Differentiation and in Age-related Bone Loss
    Article Snippet: The entire mouse Rorβ 3’UTR sequence (7279 nucleotide bases in length; Genbank Accession ), was synthesized as four approximately equal-sized gBLOCKs (Integrated DNA Technologies, Coralville, IA, USA) and assembled into the XbaI site of Luc2P-4TO using Gibson Assembly Master Mix (New England BioLabs, Ipswich, MA, USA) to produce Rorβ−3’UTR-Luc. .. Twenty-four hours later, protein and luciferase assays were performed as previously described. The data are presented as luciferase activity per μg protein and normalized to the negative control miRNA mimic.

    Infection:

    Article Title: Engineering proteins for allosteric control by light or ligands
    Article Snippet: Plasmid vectors for transient mammalian cell expression: e.g., pTriex (Addgene Plasmid # 66110), pcDNA (Addgene Plasmid # 52535) Plasmid vectors for viral infection: e.g., pBabe (Addgene Plasmid #91876), pLenti (Addgene Plasmid # 39481), FUW (Addgene Plasmid # 84008) Q5 Hot Start High-Fidelity 2X Master Mix (New England Biolabs, cat. no. M0494S) Pfu Turbo enzyme (Agilent, cat. no. 600250) CRITICAL A high fidelity DNA polymerase is required when amplifying long pieces of DNA using PCR. .. Alternatively, Gibson assembly mix (New England Biolabs, cat. no. E2611S) can be used for isothermal assembly of inserted and host plasmid DNA fragments.

    Expressing:

    Article Title: Chromobodies to Quantify Changes of Endogenous Protein Concentration in Living Cells
    Article Snippet: Paragraph title: Expression Constructs ... Fragments were assembled using the Gibson-Assembly Master Mix (New England Biolabs GmbH, Frankfurt, Germany) according to the manufacturer's protocol.

    Article Title: Engineering proteins for allosteric control by light or ligands
    Article Snippet: Plasmid vectors for transient mammalian cell expression: e.g., pTriex (Addgene Plasmid # 66110), pcDNA (Addgene Plasmid # 52535) Plasmid vectors for viral infection: e.g., pBabe (Addgene Plasmid #91876), pLenti (Addgene Plasmid # 39481), FUW (Addgene Plasmid # 84008) Q5 Hot Start High-Fidelity 2X Master Mix (New England Biolabs, cat. no. M0494S) Pfu Turbo enzyme (Agilent, cat. no. 600250) CRITICAL A high fidelity DNA polymerase is required when amplifying long pieces of DNA using PCR. .. Alternatively, Gibson assembly mix (New England Biolabs, cat. no. E2611S) can be used for isothermal assembly of inserted and host plasmid DNA fragments.

    Modification:

    Article Title: Reversibly immortalized human umbilical cord-derived mesenchymal stem cells (UC-MSCs) are responsive to BMP9-induced osteogenic and adipogenic differentiation
    Article Snippet: All cells were maintained at 37°C with 5% CO2 in Dulbecco’s modified eagle medium (DMEM) containing 10% fetal bovine serum (Gemini Bio Products, West Sacramento, CA), 2mM L-glutamine, 100 U/ml penicillin and 100 μg/ml streptomycin , . .. All restriction enzymes used for cloning, the Phusion High-Fidelity PCR kit and the Gibson Assembly Master Mix were from New England Biolabs (Ipswich, MA, USA).

    Article Title: CRISPR Activation Screens Systematically Identify Factors that Drive Neuronal Fate and Reprogramming
    Article Snippet: .. The Tubb3-IRES-hCD8 vector (pSLQ1760) was assembled with three fragments (5’ homologous arm of Tubb3, IRES-hCD8, and 3’ homologous arm of Tubb3) and a modified pUC19 backbone vector by using Gibson Assembly Master Mix (New England Biolabs). .. Both 5’ and 3’ homology arms were PCR amplified from the genomic DNA extracted from mES cells with Herculase II Fusion DNA polymerase (Agilent).

    Derivative Assay:

    Article Title: Reversibly immortalized human umbilical cord-derived mesenchymal stem cells (UC-MSCs) are responsive to BMP9-induced osteogenic and adipogenic differentiation
    Article Snippet: The 293pTP and RAPA cells were derived from HEK-293 cells as described , . .. All restriction enzymes used for cloning, the Phusion High-Fidelity PCR kit and the Gibson Assembly Master Mix were from New England Biolabs (Ipswich, MA, USA).

    Electroporation:

    Article Title: The Essential Role of Hypermutation in Rapid Adaptation to Antibiotic Stress
    Article Snippet: Allelic exchange vectors ( ) were made using the Gibson Assembly master mix (New England BioLabs) with primers listed in . .. Electroporation of the constructed plasmid into PAO1 was done at room temperature using a 2-mm-gap electroporation cuvette at 2.2 kV, as described previously ( ).

    Transfection:

    Article Title: miR-219a-5p Regulates Rorβ During Osteoblast Differentiation and in Age-related Bone Loss
    Article Snippet: The entire mouse Rorβ 3’UTR sequence (7279 nucleotide bases in length; Genbank Accession ), was synthesized as four approximately equal-sized gBLOCKs (Integrated DNA Technologies, Coralville, IA, USA) and assembled into the XbaI site of Luc2P-4TO using Gibson Assembly Master Mix (New England BioLabs, Ipswich, MA, USA) to produce Rorβ−3’UTR-Luc. .. To assess the effects of miR-219a-5p on the Rorβ 3’UTR, 100 ng of Rorβ−3’UTR-Luc were cotransfected with 100nM of either negative control or miR-219a-5p miRNA mimics (Qiagen) into primary mouse CalOBs ( n = 6) at approximately 70% confluence using DharmaFECT DUO transfection reagent (Dharmacon), a reagent specifically made for cotransfection of DNA and miRNAs.

    Cell Culture:

    Article Title: Reversibly immortalized human umbilical cord-derived mesenchymal stem cells (UC-MSCs) are responsive to BMP9-induced osteogenic and adipogenic differentiation
    Article Snippet: Paragraph title: 2.1. Cell Culture, Enzymes and Chemicals ... All restriction enzymes used for cloning, the Phusion High-Fidelity PCR kit and the Gibson Assembly Master Mix were from New England Biolabs (Ipswich, MA, USA).

    Generated:

    Article Title: Chromobodies to Quantify Changes of Endogenous Protein Concentration in Living Cells
    Article Snippet: The expression construct coding for Ubiquitin-Methionine-BC1-TagGFP2 (hereafter referred as Ub-M-BC1-TagGFP2, B ) was generated by Gibson assembly cloning ( ) of the following four fragments: fragment1 - pEGFP-N1 vector backbone digested with NheI and XbaI, fragment 2 - ubiquitin amplified from Ub-M-eGFP, a gift from Nico Dantuma (plasmid # 11938, Addgene, Cambridge, MA) , with the primer set ubiquitin-for and ubiquitin-rev, fragment 3 - BC1-NB amplified from the BC1-CB expression construct described in ( ) with the primer set BC1-for and BC1-rev, fragment 4 - TagGFP2 amplified from the BC1-CB plasmid described in ( ) with the primer set TagGFP2-for and TagGFP2-rev. .. Fragments were assembled using the Gibson-Assembly Master Mix (New England Biolabs GmbH, Frankfurt, Germany) according to the manufacturer's protocol.

    Article Title: Chromobodies to Quantify Changes of Endogenous Protein Concentration in Living Cells
    Article Snippet: Ub-M-BC1-TagGFP2 comprising a (G4 S)2 linker between BC1-NB and TagGFP2 was generated by Gibson assembly as described, with the following exception: TagGFP2 was amplified using the primer set TagGFP2–2GS-for and TagGFP2-rev. .. Fragments were assembled using the Gibson-Assembly Master Mix (New England Biolabs).

    Sequencing:

    Article Title: miR-219a-5p Regulates Rorβ During Osteoblast Differentiation and in Age-related Bone Loss
    Article Snippet: .. The entire mouse Rorβ 3’UTR sequence (7279 nucleotide bases in length; Genbank Accession ), was synthesized as four approximately equal-sized gBLOCKs (Integrated DNA Technologies, Coralville, IA, USA) and assembled into the XbaI site of Luc2P-4TO using Gibson Assembly Master Mix (New England BioLabs, Ipswich, MA, USA) to produce Rorβ−3’UTR-Luc. .. To assess the effects of miR-219a-5p on the Rorβ 3’UTR, 100 ng of Rorβ−3’UTR-Luc were cotransfected with 100nM of either negative control or miR-219a-5p miRNA mimics (Qiagen) into primary mouse CalOBs ( n = 6) at approximately 70% confluence using DharmaFECT DUO transfection reagent (Dharmacon), a reagent specifically made for cotransfection of DNA and miRNAs.

    Article Title: CRISPR Activation Screens Systematically Identify Factors that Drive Neuronal Fate and Reprogramming
    Article Snippet: The pX330-derived pSLQ1654 encoding the nuclease Cas9 and an optimized sgRNA sequence was linearized by a BbsI digest and gel purified. .. The Tubb3-IRES-hCD8 vector (pSLQ1760) was assembled with three fragments (5’ homologous arm of Tubb3, IRES-hCD8, and 3’ homologous arm of Tubb3) and a modified pUC19 backbone vector by using Gibson Assembly Master Mix (New England Biolabs).

    Molecular Cloning:

    Article Title: Chromobodies to Quantify Changes of Endogenous Protein Concentration in Living Cells
    Article Snippet: For molecular cloning of the Ub-M-BC1-TagGFP2 construct containing the longer linker (G4 S)4 Gibson assembly of the following five fragments was performed: fragment 1 - pEGFP-N1 vector backbone, and fragment 2 - ubiquitin were amplified as described above. .. Fragments were assembled using the Gibson-Assembly Master Mix (New England Biolabs).

    Article Title: Engineering proteins for allosteric control by light or ligands
    Article Snippet: Paragraph title: Molecular cloning ... Alternatively, Gibson assembly mix (New England Biolabs, cat. no. E2611S) can be used for isothermal assembly of inserted and host plasmid DNA fragments.

    Mutagenesis:

    Article Title: The Essential Role of Hypermutation in Rapid Adaptation to Antibiotic Stress
    Article Snippet: Allelic exchange vectors ( ) were made using the Gibson Assembly master mix (New England BioLabs) with primers listed in . .. Mutant pmrB alleles were amplified from the respective bioreactor-derived endpoint isolate containing the mutation.

    Article Title: Chromobodies to Quantify Changes of Endogenous Protein Concentration in Living Cells
    Article Snippet: Fragments were assembled using the Gibson-Assembly Master Mix (New England Biolabs GmbH, Frankfurt, Germany) according to the manufacturer's protocol. .. The corresponding N-terminal mutants of the Ub-M-BC1-TagGFP2 expression construct were generated by site-directed mutagenesis of the Ub-M-BC1-TagGFP2 plasmid with the primer pair NB-N-term-X-mut-for (X represents the corresponding N-terminal amino acid) and NB-N-term-rev using the Q5 Site-Directed Mutagenesis Kit (New England Biolabs) according to the manufacturer's protocol.

    Article Title: Chromobodies to Quantify Changes of Endogenous Protein Concentration in Living Cells
    Article Snippet: The noncleavable mutant UbG76V -M-BC1-TagGFP2 was produced by site-directed mutagenesis using the primer set BC1-for and Ub-G76V-mut-rev. .. Fragments were assembled using the Gibson-Assembly Master Mix (New England Biolabs).

    Purification:

    Article Title: CRISPR Activation Screens Systematically Identify Factors that Drive Neuronal Fate and Reprogramming
    Article Snippet: The pX330-derived pSLQ1654 encoding the nuclease Cas9 and an optimized sgRNA sequence was linearized by a BbsI digest and gel purified. .. The Tubb3-IRES-hCD8 vector (pSLQ1760) was assembled with three fragments (5’ homologous arm of Tubb3, IRES-hCD8, and 3’ homologous arm of Tubb3) and a modified pUC19 backbone vector by using Gibson Assembly Master Mix (New England Biolabs).

    Polymerase Chain Reaction:

    Article Title: Reversibly immortalized human umbilical cord-derived mesenchymal stem cells (UC-MSCs) are responsive to BMP9-induced osteogenic and adipogenic differentiation
    Article Snippet: .. All restriction enzymes used for cloning, the Phusion High-Fidelity PCR kit and the Gibson Assembly Master Mix were from New England Biolabs (Ipswich, MA, USA). ..

    Article Title: Broadened Glycosylation Patterning of Heterologously Produced Erythromycin
    Article Snippet: .. The chaperonin plasmid pGro7 was obtained from Takara (Madison, WI), and restriction endonucleases, T4 DNA ligase, Phusion High-Fidelity PCR Master Mix, and Gibson Assembly Master Mix were purchased from NEB (Ipswich, MA). provides a summary of the plasmids and strains in this work. ..

    Article Title: CRISPR Activation Screens Systematically Identify Factors that Drive Neuronal Fate and Reprogramming
    Article Snippet: The Tubb3-IRES-hCD8 vector (pSLQ1760) was assembled with three fragments (5’ homologous arm of Tubb3, IRES-hCD8, and 3’ homologous arm of Tubb3) and a modified pUC19 backbone vector by using Gibson Assembly Master Mix (New England Biolabs). .. Both 5’ and 3’ homology arms were PCR amplified from the genomic DNA extracted from mES cells with Herculase II Fusion DNA polymerase (Agilent).

    Article Title: Engineering proteins for allosteric control by light or ligands
    Article Snippet: Plasmid vectors for transient mammalian cell expression: e.g., pTriex (Addgene Plasmid # 66110), pcDNA (Addgene Plasmid # 52535) Plasmid vectors for viral infection: e.g., pBabe (Addgene Plasmid #91876), pLenti (Addgene Plasmid # 39481), FUW (Addgene Plasmid # 84008) Q5 Hot Start High-Fidelity 2X Master Mix (New England Biolabs, cat. no. M0494S) Pfu Turbo enzyme (Agilent, cat. no. 600250) CRITICAL A high fidelity DNA polymerase is required when amplifying long pieces of DNA using PCR. .. Alternatively, Gibson assembly mix (New England Biolabs, cat. no. E2611S) can be used for isothermal assembly of inserted and host plasmid DNA fragments.

    Cotransfection:

    Article Title: miR-219a-5p Regulates Rorβ During Osteoblast Differentiation and in Age-related Bone Loss
    Article Snippet: The entire mouse Rorβ 3’UTR sequence (7279 nucleotide bases in length; Genbank Accession ), was synthesized as four approximately equal-sized gBLOCKs (Integrated DNA Technologies, Coralville, IA, USA) and assembled into the XbaI site of Luc2P-4TO using Gibson Assembly Master Mix (New England BioLabs, Ipswich, MA, USA) to produce Rorβ−3’UTR-Luc. .. To assess the effects of miR-219a-5p on the Rorβ 3’UTR, 100 ng of Rorβ−3’UTR-Luc were cotransfected with 100nM of either negative control or miR-219a-5p miRNA mimics (Qiagen) into primary mouse CalOBs ( n = 6) at approximately 70% confluence using DharmaFECT DUO transfection reagent (Dharmacon), a reagent specifically made for cotransfection of DNA and miRNAs.

    Plasmid Preparation:

    Article Title: miR-219a-5p Regulates Rorβ During Osteoblast Differentiation and in Age-related Bone Loss
    Article Snippet: The luc2P fragment was subcloned into the HindIII/XbaI sites of pcDNA/4TO (Promega) to make the 3’UTR reporter vector, Luc2P-4TO. .. The entire mouse Rorβ 3’UTR sequence (7279 nucleotide bases in length; Genbank Accession ), was synthesized as four approximately equal-sized gBLOCKs (Integrated DNA Technologies, Coralville, IA, USA) and assembled into the XbaI site of Luc2P-4TO using Gibson Assembly Master Mix (New England BioLabs, Ipswich, MA, USA) to produce Rorβ−3’UTR-Luc.

    Article Title: The Essential Role of Hypermutation in Rapid Adaptation to Antibiotic Stress
    Article Snippet: Allelic exchange vectors ( ) were made using the Gibson Assembly master mix (New England BioLabs) with primers listed in . .. Electroporation of the constructed plasmid into PAO1 was done at room temperature using a 2-mm-gap electroporation cuvette at 2.2 kV, as described previously ( ).

    Article Title: Chromobodies to Quantify Changes of Endogenous Protein Concentration in Living Cells
    Article Snippet: The expression construct coding for Ubiquitin-Methionine-BC1-TagGFP2 (hereafter referred as Ub-M-BC1-TagGFP2, B ) was generated by Gibson assembly cloning ( ) of the following four fragments: fragment1 - pEGFP-N1 vector backbone digested with NheI and XbaI, fragment 2 - ubiquitin amplified from Ub-M-eGFP, a gift from Nico Dantuma (plasmid # 11938, Addgene, Cambridge, MA) , with the primer set ubiquitin-for and ubiquitin-rev, fragment 3 - BC1-NB amplified from the BC1-CB expression construct described in ( ) with the primer set BC1-for and BC1-rev, fragment 4 - TagGFP2 amplified from the BC1-CB plasmid described in ( ) with the primer set TagGFP2-for and TagGFP2-rev. .. Fragments were assembled using the Gibson-Assembly Master Mix (New England Biolabs GmbH, Frankfurt, Germany) according to the manufacturer's protocol.

    Article Title: Broadened Glycosylation Patterning of Heterologously Produced Erythromycin
    Article Snippet: .. The chaperonin plasmid pGro7 was obtained from Takara (Madison, WI), and restriction endonucleases, T4 DNA ligase, Phusion High-Fidelity PCR Master Mix, and Gibson Assembly Master Mix were purchased from NEB (Ipswich, MA). provides a summary of the plasmids and strains in this work. ..

    Article Title: CRISPR Activation Screens Systematically Identify Factors that Drive Neuronal Fate and Reprogramming
    Article Snippet: .. The Tubb3-IRES-hCD8 vector (pSLQ1760) was assembled with three fragments (5’ homologous arm of Tubb3, IRES-hCD8, and 3’ homologous arm of Tubb3) and a modified pUC19 backbone vector by using Gibson Assembly Master Mix (New England Biolabs). .. Both 5’ and 3’ homology arms were PCR amplified from the genomic DNA extracted from mES cells with Herculase II Fusion DNA polymerase (Agilent).

    Article Title: Chromobodies to Quantify Changes of Endogenous Protein Concentration in Living Cells
    Article Snippet: Fragment 3 - BC1-NB was amplified from the BC1-CB expression construct with the primer set BC1-for and BC1–4GS-rev, fragment 4 - TagGFP2 was amplified from the BC1-TagGFP2 plasmid with the primer set TagGFP2–4GS-for and TagGFP2-rev, fragment 5 - (G4 S)4 -linker was produced by annealing of the two single stranded oligonucleotides 4GS-linker-oligo1 and 4GS-linker-oligo2 with complementary sequences. .. Fragments were assembled using the Gibson-Assembly Master Mix (New England Biolabs).

    Article Title: Engineering proteins for allosteric control by light or ligands
    Article Snippet: .. Alternatively, Gibson assembly mix (New England Biolabs, cat. no. E2611S) can be used for isothermal assembly of inserted and host plasmid DNA fragments. .. DNA amplification kit (Fischer Scientific, cat. no. FERK0503 and Qiagen, cat. no. 12643) CAUTION Buffers in the kit can cause serious eye and skin irritation.

    Negative Control:

    Article Title: miR-219a-5p Regulates Rorβ During Osteoblast Differentiation and in Age-related Bone Loss
    Article Snippet: The entire mouse Rorβ 3’UTR sequence (7279 nucleotide bases in length; Genbank Accession ), was synthesized as four approximately equal-sized gBLOCKs (Integrated DNA Technologies, Coralville, IA, USA) and assembled into the XbaI site of Luc2P-4TO using Gibson Assembly Master Mix (New England BioLabs, Ipswich, MA, USA) to produce Rorβ−3’UTR-Luc. .. To assess the effects of miR-219a-5p on the Rorβ 3’UTR, 100 ng of Rorβ−3’UTR-Luc were cotransfected with 100nM of either negative control or miR-219a-5p miRNA mimics (Qiagen) into primary mouse CalOBs ( n = 6) at approximately 70% confluence using DharmaFECT DUO transfection reagent (Dharmacon), a reagent specifically made for cotransfection of DNA and miRNAs.

    Produced:

    Article Title: Chromobodies to Quantify Changes of Endogenous Protein Concentration in Living Cells
    Article Snippet: Fragments were assembled using the Gibson-Assembly Master Mix (New England Biolabs GmbH, Frankfurt, Germany) according to the manufacturer's protocol. .. The noncleavable mutant UbG76V -M-BC1-TagGFP2 was produced by site-directed mutagenesis using the primer set BC1-for and Ub-G76V-mut-rev.

    Article Title: Chromobodies to Quantify Changes of Endogenous Protein Concentration in Living Cells
    Article Snippet: Fragment 3 - BC1-NB was amplified from the BC1-CB expression construct with the primer set BC1-for and BC1–4GS-rev, fragment 4 - TagGFP2 was amplified from the BC1-TagGFP2 plasmid with the primer set TagGFP2–4GS-for and TagGFP2-rev, fragment 5 - (G4 S)4 -linker was produced by annealing of the two single stranded oligonucleotides 4GS-linker-oligo1 and 4GS-linker-oligo2 with complementary sequences. .. Fragments were assembled using the Gibson-Assembly Master Mix (New England Biolabs).

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    New England Biolabs gibson assembly master mix
    Gibson Assembly Master Mix, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 90/100, based on 454 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 90 stars, based on 454 article reviews
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