dh5α  (New England Biolabs)


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    Structured Review

    New England Biolabs dh5α
    Dh5α, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 92/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dh5α/product/New England Biolabs
    Average 92 stars, based on 9 article reviews
    Price from $9.99 to $1999.99
    dh5α - by Bioz Stars, 2020-10
    92/100 stars

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    Related Articles

    Clone Assay:

    Article Title: Diverse protein assembly driven by metal and chelating amino acids with selectivity and tunability
    Article Snippet: .. For gene cloning and heterologous protein expression, DH5α and BL21 (DE3) E. coli strains (NEB) were used, respectively. .. For protein expression, plasmids were transformed to BL21(DE3) E. coli competent cells (NEB), containing pEVOL plasmid for the incorporation of Bpy-Ala .

    Affinity Chromatography:

    Article Title: Dissection of the Functional and Structural Domains of Phosphorelay Histidine Kinase A of Bacillus subtilis †
    Article Snippet: .. The MBP fusion proteins were purified from DH5α by amylose affinity chromatography, according to the specifications of the manufacturer (New England Biolabs). .. Briefly, cells were disrupted by sonication (8 times for 15 s each time) in sonication buffer (SB) (25 mM Tris [pH 8.0], 1 mM EDTA, 1 mM dithiothreitol [DTT], 10 mM KCl, 5 mM MgCl2 , 1 mM phenylmethylsulfonyl fluoride [PMSF]), and then debris and membranes were removed by centrifugation.

    Construct:

    Article Title: Unique nucleotide sequence (UNS)-guided assembly of repetitive DNA parts for synthetic biology applications
    Article Snippet: .. We note that E. coli with mutations in recA should be used (e.g. DH5α, TOP10 or Mach1), as using strains with wild-type recA (e.g. NEB TURBO) considerably increases the proportion of incorrectly-assembled constructs. .. The successful assembly of the desired products can typically be validated by culturing 2–3 colonies, mini-prepping their DNA, and performing analytical restriction digestions to confirm that the isolated plasmids are of the correct size.

    Purification:

    Article Title: The NIMA-family kinase Nek6 phosphorylates the kinesin Eg5 at a novel site necessary for mitotic spindle formation
    Article Snippet: .. MalBP-Eg5 fusion protein expression in DH5α was induced with 0.4mM IPTG for 5h at 37°C and purified using amylose resin according to the manufacturers instructions (New England Biolabs, Ipswich, MA, USA). .. GST-Eg5 was expressed in 293T cells by transient transfection.

    Article Title: Dissection of the Functional and Structural Domains of Phosphorelay Histidine Kinase A of Bacillus subtilis †
    Article Snippet: .. The MBP fusion proteins were purified from DH5α by amylose affinity chromatography, according to the specifications of the manufacturer (New England Biolabs). .. Briefly, cells were disrupted by sonication (8 times for 15 s each time) in sonication buffer (SB) (25 mM Tris [pH 8.0], 1 mM EDTA, 1 mM dithiothreitol [DTT], 10 mM KCl, 5 mM MgCl2 , 1 mM phenylmethylsulfonyl fluoride [PMSF]), and then debris and membranes were removed by centrifugation.

    Article Title: Highly efficient ‘hit-and-run’ genome editing with unconcentrated lentivectors carrying Vpr.Prot.Cas9 protein produced from RRE-containing transcripts
    Article Snippet: .. Plasmids were amplified in DH5α or NEB Stable competent Escherichia coli (New England Biolabs) and purified using a Qiagen Plasmid Midi kit (Qiagen). .. Lentivector production and transduction To produce lentivectors containing both Cas9 protein and U6-sgRNA template five plasmids (a total amount of 4 μg) were co-transfected to exponentially growing HEK293T cells seeded in six-well plates (ATCC CRL-3216; ∼80% confluent) using Fugene HD (Promega).

    Amplification:

    Article Title: Highly efficient ‘hit-and-run’ genome editing with unconcentrated lentivectors carrying Vpr.Prot.Cas9 protein produced from RRE-containing transcripts
    Article Snippet: .. Plasmids were amplified in DH5α or NEB Stable competent Escherichia coli (New England Biolabs) and purified using a Qiagen Plasmid Midi kit (Qiagen). .. Lentivector production and transduction To produce lentivectors containing both Cas9 protein and U6-sgRNA template five plasmids (a total amount of 4 μg) were co-transfected to exponentially growing HEK293T cells seeded in six-well plates (ATCC CRL-3216; ∼80% confluent) using Fugene HD (Promega).

    Expressing:

    Article Title: The NIMA-family kinase Nek6 phosphorylates the kinesin Eg5 at a novel site necessary for mitotic spindle formation
    Article Snippet: .. MalBP-Eg5 fusion protein expression in DH5α was induced with 0.4mM IPTG for 5h at 37°C and purified using amylose resin according to the manufacturers instructions (New England Biolabs, Ipswich, MA, USA). .. GST-Eg5 was expressed in 293T cells by transient transfection.

    Article Title: Diverse protein assembly driven by metal and chelating amino acids with selectivity and tunability
    Article Snippet: .. For gene cloning and heterologous protein expression, DH5α and BL21 (DE3) E. coli strains (NEB) were used, respectively. .. For protein expression, plasmids were transformed to BL21(DE3) E. coli competent cells (NEB), containing pEVOL plasmid for the incorporation of Bpy-Ala .

    Transformation Assay:

    Article Title: O-GlcNAc transferase associates with the MCM2–7 complex and its silencing destabilizes MCM–MCM interactions
    Article Snippet: .. Prior to transformation in DH5α, template was digested for 2 h at 37 °C by 1 U of DpnI (NEB). .. Cell culture, transfection and cell cycle synchronization MCF7 and MDA-MB-231 breast cancer cell lines, and HEK 293T cells were routinely grown at 37 °C in a humidified atmosphere enriched with 5% CO2 in Dulbecco’s modified Eagle’s medium (DMEM) (Lonza, Basel, Switzerland) containing high glucose (4.5 g/L) and glutamine, and supplemented with 10% foetal calf serum (FCS) (Lonza) (complete medium).

    Plasmid Preparation:

    Article Title: Highly efficient ‘hit-and-run’ genome editing with unconcentrated lentivectors carrying Vpr.Prot.Cas9 protein produced from RRE-containing transcripts
    Article Snippet: .. Plasmids were amplified in DH5α or NEB Stable competent Escherichia coli (New England Biolabs) and purified using a Qiagen Plasmid Midi kit (Qiagen). .. Lentivector production and transduction To produce lentivectors containing both Cas9 protein and U6-sgRNA template five plasmids (a total amount of 4 μg) were co-transfected to exponentially growing HEK293T cells seeded in six-well plates (ATCC CRL-3216; ∼80% confluent) using Fugene HD (Promega).

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  • 92
    New England Biolabs dh5α strain
    MAIT cells respond to and control CREC. (A) Expression of CD107a, GrzB, IFNγ, TNF, and IL-17A in MAIT cells stimulated for 24 h with E . coli strains <t>DH5α</t> ( n = 16), EC120S ( n = 7), and the carbapenem-resistant strains EC234 ( n = 16), EC241 ( n = 5), EC362 ( n = 16), and EC385 ( n = 5). Unstimulated, n = 16. (B) MR1-dependency of effector protein and cytokine production by MAIT cells stimulated with indicated strains. MR1-dependency was calculated as previously described [ 2 ] ( n = 7). (C) Representative flow cytometry plots of Casp3 activation and apoptosis in HeLa cells alone, HeLa cells infected with EC241, or co-cultured with MAIT cells with or without EC241 for 3 h. (D) Apoptosis of HeLa cells alone or co-cultured with MAIT cells for 3 h in the presence of 5-OP-RU, EC120S, EC234, EC241, or EC362 ( n = 5). (E, F) Casp3 activation (E) and bacterial loads (F) in HeLa cells infected with EC241 co-cultured with MAIT cells in the presence of anti-MR1 antibody or isotype control ( n = 5–8 for EC241-infected HeLa cells+anti-MR1 mAb, n = 13–25 others). Data presented as bar graphs with error bars represent the mean and standard error. Box and whisker plots show median, the 10th to 90th percentile, and the interquartile range. Statistical significance was determined using the Kruskal-Wallis ANOVA (A) or the Friedman test (B, D) followed by Dunn’s multiple comparison test, or mixed-effects analysis followed by Dunnett’s multiple comparison test (E, F). **** p
    Dh5α Strain, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 92/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dh5α strain/product/New England Biolabs
    Average 92 stars, based on 6 article reviews
    Price from $9.99 to $1999.99
    dh5α strain - by Bioz Stars, 2020-10
    92/100 stars
      Buy from Supplier

    92
    New England Biolabs dh5α cells
    TEM confirmation of horizontal gene transfer among E. coli expressing pSF-OXB15-p450camfusion and P. putida . A. E. coli <t>DH5α</t> (E) connected to P. putida (P) via conjugative pili. B. Mating-pair bridge between P. putida and E. coli DH5α. C. E. coli DH5α with conjugative pili. D. Nanotube connecting P. putida and E.coli DH5α.
    Dh5α Cells, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 92/100, based on 28 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/dh5α cells/product/New England Biolabs
    Average 92 stars, based on 28 article reviews
    Price from $9.99 to $1999.99
    dh5α cells - by Bioz Stars, 2020-10
    92/100 stars
      Buy from Supplier

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    MAIT cells respond to and control CREC. (A) Expression of CD107a, GrzB, IFNγ, TNF, and IL-17A in MAIT cells stimulated for 24 h with E . coli strains DH5α ( n = 16), EC120S ( n = 7), and the carbapenem-resistant strains EC234 ( n = 16), EC241 ( n = 5), EC362 ( n = 16), and EC385 ( n = 5). Unstimulated, n = 16. (B) MR1-dependency of effector protein and cytokine production by MAIT cells stimulated with indicated strains. MR1-dependency was calculated as previously described [ 2 ] ( n = 7). (C) Representative flow cytometry plots of Casp3 activation and apoptosis in HeLa cells alone, HeLa cells infected with EC241, or co-cultured with MAIT cells with or without EC241 for 3 h. (D) Apoptosis of HeLa cells alone or co-cultured with MAIT cells for 3 h in the presence of 5-OP-RU, EC120S, EC234, EC241, or EC362 ( n = 5). (E, F) Casp3 activation (E) and bacterial loads (F) in HeLa cells infected with EC241 co-cultured with MAIT cells in the presence of anti-MR1 antibody or isotype control ( n = 5–8 for EC241-infected HeLa cells+anti-MR1 mAb, n = 13–25 others). Data presented as bar graphs with error bars represent the mean and standard error. Box and whisker plots show median, the 10th to 90th percentile, and the interquartile range. Statistical significance was determined using the Kruskal-Wallis ANOVA (A) or the Friedman test (B, D) followed by Dunn’s multiple comparison test, or mixed-effects analysis followed by Dunnett’s multiple comparison test (E, F). **** p

    Journal: PLoS Biology

    Article Title: Human MAIT cell cytolytic effector proteins synergize to overcome carbapenem resistance in Escherichia coli

    doi: 10.1371/journal.pbio.3000644

    Figure Lengend Snippet: MAIT cells respond to and control CREC. (A) Expression of CD107a, GrzB, IFNγ, TNF, and IL-17A in MAIT cells stimulated for 24 h with E . coli strains DH5α ( n = 16), EC120S ( n = 7), and the carbapenem-resistant strains EC234 ( n = 16), EC241 ( n = 5), EC362 ( n = 16), and EC385 ( n = 5). Unstimulated, n = 16. (B) MR1-dependency of effector protein and cytokine production by MAIT cells stimulated with indicated strains. MR1-dependency was calculated as previously described [ 2 ] ( n = 7). (C) Representative flow cytometry plots of Casp3 activation and apoptosis in HeLa cells alone, HeLa cells infected with EC241, or co-cultured with MAIT cells with or without EC241 for 3 h. (D) Apoptosis of HeLa cells alone or co-cultured with MAIT cells for 3 h in the presence of 5-OP-RU, EC120S, EC234, EC241, or EC362 ( n = 5). (E, F) Casp3 activation (E) and bacterial loads (F) in HeLa cells infected with EC241 co-cultured with MAIT cells in the presence of anti-MR1 antibody or isotype control ( n = 5–8 for EC241-infected HeLa cells+anti-MR1 mAb, n = 13–25 others). Data presented as bar graphs with error bars represent the mean and standard error. Box and whisker plots show median, the 10th to 90th percentile, and the interquartile range. Statistical significance was determined using the Kruskal-Wallis ANOVA (A) or the Friedman test (B, D) followed by Dunn’s multiple comparison test, or mixed-effects analysis followed by Dunnett’s multiple comparison test (E, F). **** p

    Article Snippet: Bacterial cultures The E . coli strains 1100–2 and BSV18 were obtained from the Coli Genetic Stock Center, Yale University; the DH5α strain was obtained from New England Biolab.

    Techniques: Expressing, Flow Cytometry, Activation Assay, Infection, Cell Culture, Whisker Assay

    The biological parts utilised for combinatorial construction of a lycopene biosynthetic pathway. a The carotenoid lycopene is produced in E. coli through a heterologous biosynthetic pathway composed of three enzymes (CrtE. CrtB and CrtI). The heterologous proteins use FPP, which is a product of MEP pathway, as a precursor. b : Table S1). The pathway library was assembled and expressed in E. coli DH5α to test lycopene titres. The relative effects of the different design factors had on the titres level of lycopene were modelled using JMP. The model identified biological parts which effected titre levels, this can help aid future designing of the lycopene biosynthetic pathway.

    Journal: Journal of Biological Engineering

    Article Title: Utilising datasheets for the informed automated design and build of a synthetic metabolic pathway

    doi: 10.1186/s13036-019-0141-z

    Figure Lengend Snippet: The biological parts utilised for combinatorial construction of a lycopene biosynthetic pathway. a The carotenoid lycopene is produced in E. coli through a heterologous biosynthetic pathway composed of three enzymes (CrtE. CrtB and CrtI). The heterologous proteins use FPP, which is a product of MEP pathway, as a precursor. b : Table S1). The pathway library was assembled and expressed in E. coli DH5α to test lycopene titres. The relative effects of the different design factors had on the titres level of lycopene were modelled using JMP. The model identified biological parts which effected titre levels, this can help aid future designing of the lycopene biosynthetic pathway.

    Article Snippet: The E. coli strain DH5α (NEB, USA) was used for all DNA cloning.

    Techniques: Produced

    The biological parts utilised for combinatorial construction of a lycopene biosynthetic pathway. a The carotenoid lycopene is produced in E. coli through a heterologous biosynthetic pathway composed of three enzymes (CrtE. CrtB and CrtI). The heterologous proteins use FPP, which is a product of MEP pathway, as a precursor. b A complete combinatorial library totalling 810 pathway configurations can be designed by varying the promoter and RBS parts and by varying the order of pathway genes and RBS parts. Through the application of statistical modelling by DoE software, the designed library was reduced to 88 representative constructs, A table displaying part information of each construct is displayed in the supplementary information ( Additional file 1 : Table S1). The pathway library was assembled and expressed in E. coli DH5α to test lycopene titres. The relative effects of the different design factors had on the titres level of lycopene were modelled using JMP. The model identified biological parts which effected titre levels, this can help aid future designing of the lycopene biosynthetic pathway.

    Journal: Journal of Biological Engineering

    Article Title: Utilising datasheets for the informed automated design and build of a synthetic metabolic pathway

    doi: 10.1186/s13036-019-0141-z

    Figure Lengend Snippet: The biological parts utilised for combinatorial construction of a lycopene biosynthetic pathway. a The carotenoid lycopene is produced in E. coli through a heterologous biosynthetic pathway composed of three enzymes (CrtE. CrtB and CrtI). The heterologous proteins use FPP, which is a product of MEP pathway, as a precursor. b A complete combinatorial library totalling 810 pathway configurations can be designed by varying the promoter and RBS parts and by varying the order of pathway genes and RBS parts. Through the application of statistical modelling by DoE software, the designed library was reduced to 88 representative constructs, A table displaying part information of each construct is displayed in the supplementary information ( Additional file 1 : Table S1). The pathway library was assembled and expressed in E. coli DH5α to test lycopene titres. The relative effects of the different design factors had on the titres level of lycopene were modelled using JMP. The model identified biological parts which effected titre levels, this can help aid future designing of the lycopene biosynthetic pathway.

    Article Snippet: The E. coli strain DH5α (NEB, USA) was used for all DNA cloning.

    Techniques: Produced, Software, Construct

    TEM confirmation of horizontal gene transfer among E. coli expressing pSF-OXB15-p450camfusion and P. putida . A. E. coli DH5α (E) connected to P. putida (P) via conjugative pili. B. Mating-pair bridge between P. putida and E. coli DH5α. C. E. coli DH5α with conjugative pili. D. Nanotube connecting P. putida and E.coli DH5α.

    Journal: bioRxiv

    Article Title: Horizontal ‘gene drives’ harness indigenous bacteria for bioremediation

    doi: 10.1101/735886

    Figure Lengend Snippet: TEM confirmation of horizontal gene transfer among E. coli expressing pSF-OXB15-p450camfusion and P. putida . A. E. coli DH5α (E) connected to P. putida (P) via conjugative pili. B. Mating-pair bridge between P. putida and E. coli DH5α. C. E. coli DH5α with conjugative pili. D. Nanotube connecting P. putida and E.coli DH5α.

    Article Snippet: To transform E.coli , we added 5 μl of ligation reaction to DH5α cells (NEB), incubated cells on ice for 30 minutes, heat-shocked cells for 60 seconds, and allowed cells to recover for five minutes on ice.

    Techniques: Transmission Electron Microscopy, Expressing

    Growth of wild-type and engineered bacteria on dodecane (A), benzo(a)pyrene (B), and crude oil (C). Wild type strains are denoted as ‘ Cupriavidus ’ and ‘ P. putida .’ Synthetic strains are denoted according to what enzyme they are engineered to express (e.g. alkB, almA). The two negative controls are a control with the carbon substrate but no cells and E. coli DH5α transformed with the vector backbone used in the experiment (pSFOXB15) but without genes inserted for hydrocarbon degradation.

    Journal: bioRxiv

    Article Title: Horizontal ‘gene drives’ harness indigenous bacteria for bioremediation

    doi: 10.1101/735886

    Figure Lengend Snippet: Growth of wild-type and engineered bacteria on dodecane (A), benzo(a)pyrene (B), and crude oil (C). Wild type strains are denoted as ‘ Cupriavidus ’ and ‘ P. putida .’ Synthetic strains are denoted according to what enzyme they are engineered to express (e.g. alkB, almA). The two negative controls are a control with the carbon substrate but no cells and E. coli DH5α transformed with the vector backbone used in the experiment (pSFOXB15) but without genes inserted for hydrocarbon degradation.

    Article Snippet: To transform E.coli , we added 5 μl of ligation reaction to DH5α cells (NEB), incubated cells on ice for 30 minutes, heat-shocked cells for 60 seconds, and allowed cells to recover for five minutes on ice.

    Techniques: Transformation Assay, Plasmid Preparation

    Expression and localization of bacterial monooxygenases and dioxygenases involved in petroleum degradation in E . coli DH5α. A. Structured Illumination Microscopy (SIM) image of E.coli DH5α expressing proteins involved in petroleum degradation (cam A,B, C and D) from the CAM plasmid in E.coli . camC (fused to mcherry) was found throughout the cell while camD (fused to gfp) localized to a microcompartment at one end of the cell. The scale bar is 5 μm. B. E. coli DH5α expressing alkB fuse to gfp were found clinging to spheres containing crude oil, mimicking a behavior seen in wild-type oil-degrading bacteria. C. EPS from E. coli DH5α expressing xylE. Gfp-tagged xylE were found around small pores (ca. 500 nm) within the EPS matrix. Figures 1B and 1C were taken using the GFP filter on a Zeiss AxioImager M1.

    Journal: bioRxiv

    Article Title: Horizontal ‘gene drives’ harness indigenous bacteria for bioremediation

    doi: 10.1101/735886

    Figure Lengend Snippet: Expression and localization of bacterial monooxygenases and dioxygenases involved in petroleum degradation in E . coli DH5α. A. Structured Illumination Microscopy (SIM) image of E.coli DH5α expressing proteins involved in petroleum degradation (cam A,B, C and D) from the CAM plasmid in E.coli . camC (fused to mcherry) was found throughout the cell while camD (fused to gfp) localized to a microcompartment at one end of the cell. The scale bar is 5 μm. B. E. coli DH5α expressing alkB fuse to gfp were found clinging to spheres containing crude oil, mimicking a behavior seen in wild-type oil-degrading bacteria. C. EPS from E. coli DH5α expressing xylE. Gfp-tagged xylE were found around small pores (ca. 500 nm) within the EPS matrix. Figures 1B and 1C were taken using the GFP filter on a Zeiss AxioImager M1.

    Article Snippet: To transform E.coli , we added 5 μl of ligation reaction to DH5α cells (NEB), incubated cells on ice for 30 minutes, heat-shocked cells for 60 seconds, and allowed cells to recover for five minutes on ice.

    Techniques: Expressing, Microscopy, Chick Chorioallantoic Membrane Assay, Plasmid Preparation

    Overview of horizontal “gene-drive” system. The diagram depicts two approaches to flooding indigenous bacterial populations with catabolic genes of interest 1) through controlled mating and re-release or 2) direct application of E.coli DH5α with the catabolic genes of interest. Step 4 is there as a check to ensure the native bacteria take up the plasmid while in a soil matrix and can be shortened/extended as needed. The final step 5 is for monitoring purposes and can be shortened/extended as needed. This protocol works with water/marine samples (substituting water for soil).

    Journal: bioRxiv

    Article Title: Horizontal ‘gene drives’ harness indigenous bacteria for bioremediation

    doi: 10.1101/735886

    Figure Lengend Snippet: Overview of horizontal “gene-drive” system. The diagram depicts two approaches to flooding indigenous bacterial populations with catabolic genes of interest 1) through controlled mating and re-release or 2) direct application of E.coli DH5α with the catabolic genes of interest. Step 4 is there as a check to ensure the native bacteria take up the plasmid while in a soil matrix and can be shortened/extended as needed. The final step 5 is for monitoring purposes and can be shortened/extended as needed. This protocol works with water/marine samples (substituting water for soil).

    Article Snippet: To transform E.coli , we added 5 μl of ligation reaction to DH5α cells (NEB), incubated cells on ice for 30 minutes, heat-shocked cells for 60 seconds, and allowed cells to recover for five minutes on ice.

    Techniques: Plasmid Preparation