e coli dh5α  (Thermo Fisher)


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    Structured Review

    Thermo Fisher e coli dh5α
    PCR amplification of E.coli <t>DH5α</t> clones Lane 2 to 7 –positive amplicons Lane1- DNA marker
    E Coli Dh5α, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 96/100, based on 121 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/e coli dh5α/product/Thermo Fisher
    Average 96 stars, based on 121 article reviews
    Price from $9.99 to $1999.99
    e coli dh5α - by Bioz Stars, 2020-01
    96/100 stars

    Images

    1) Product Images from "Cloning and sequence analysis of a partial CDS of leptospiral ligA gene in pET-32a – Escherichia coli DH5α system"

    Article Title: Cloning and sequence analysis of a partial CDS of leptospiral ligA gene in pET-32a – Escherichia coli DH5α system

    Journal: Veterinary World

    doi: 10.14202/vetworld.2018.557-561

    PCR amplification of E.coli DH5α clones Lane 2 to 7 –positive amplicons Lane1- DNA marker
    Figure Legend Snippet: PCR amplification of E.coli DH5α clones Lane 2 to 7 –positive amplicons Lane1- DNA marker

    Techniques Used: Polymerase Chain Reaction, Amplification, Clone Assay, Marker

    2) Product Images from "Cloning and sequence analysis of a partial CDS of leptospiral ligA gene in pET-32a – Escherichia coli DH5α system"

    Article Title: Cloning and sequence analysis of a partial CDS of leptospiral ligA gene in pET-32a – Escherichia coli DH5α system

    Journal: Veterinary World

    doi: 10.14202/vetworld.2018.557-561

    PCR amplification of E.coli DH5α clones Lane 2 to 7 –positive amplicons Lane1- DNA marker
    Figure Legend Snippet: PCR amplification of E.coli DH5α clones Lane 2 to 7 –positive amplicons Lane1- DNA marker

    Techniques Used: Polymerase Chain Reaction, Amplification, Clone Assay, Marker

    Related Articles

    Clone Assay:

    Article Title: RNA polymerase-induced remodelling of NusA produces a pause enhancement complex
    Article Snippet: .. DNA manipulation All cloning was carried out using E. coli DH5α (Gibco BRL; Supplementary Table S1). ..

    Article Title: The multi-catalytic compartment of propionyl-CoA synthase sequesters a toxic metabolite
    Article Snippet: .. E. coli DH5α (Thermo Scientific™) strains were used for cloning and grown in LB medium . .. For protein expression E. coli BL21-AI™ (Invitrogen) or Arctic-Express (DE3) RIL (Agilent Technologies) were grown in TB medium .

    Article Title: Feedback Inhibition of Chorismate Mutase/Prephenate Dehydrogenase (TyrA) of Escherichia coli: Generation and Characterization of Tyrosine-Insensitive Mutants
    Article Snippet: .. After gel purification, the PCR products were subcloned into pET-30 Xa/LIC (Novagen) by using ligation-independent cloning (LIC; Novagen) and transformed E. coli DH5α (Invitrogen). .. The resulting plasmids, pET30:: tyrA fbr , were analyzed by DNA sequencing with the following primers: T7_prom (5′-TAA TAC GAC TCA CTA TAG GG-3′), T7_term (5′-GCT AGT TAT TGC TCA GCG G-3′), tyrA_291fw (5′-ACT GCG TCC GGT GGT TAT CG-3′), and tyrA_913rv (5′-GGC GAA GAG AGC GCC AGA AG-3′).

    Article Title: Kinetic Properties of Four Plasmid-Mediated AmpC ?-Lactamases
    Article Snippet: .. E. coli DH5α (Gibco-BRL-Life Technologies, Eragny, France) and E. coli BL21(DE3) (Novagen Inc., Madison, WI) were used as recipients for cloning and expression experiments, respectively. .. The plasmid pGEM-T-easy from Promega (Madison, WI) was used to clone the PCR products and pET26b(+) (Novagen, Inc.) was used to produce the different plasmid-encoded class C β-lactamases.

    Article Title: A Novel Epimerase That Converts GlcNAc-P-P-undecaprenol to GalNAc-P-P-undecaprenol in Escherichia coli O157 *
    Article Snippet: .. E. coli DH5α (Invitrogen) was used as the host for cloning experiments and for protein glycosylation analysis. ..

    Article Title: Aggregating sequences that occur in many proteins constitute weak spots of bacterial proteostasis
    Article Snippet: .. Escherichia coli DH5α (Thermo Fisher Scientific) was used for cloning and plasmid amplification. .. For selection of antibiotic resistance colonies, E . coli carrying plasmids was grown in LB medium supplemented with the relevant antibiotic.

    Centrifugation:

    Article Title: Enhanced performance of the microalga Chlorella sorokiniana remotely induced by the plant growth-promoting bacteria Azospirillum brasilense and Bacillus pumilus
    Article Snippet: In experiments that measured the potential effect of CO2 , Escherichia coli DH5α (Invitrogen, Carlsbad, CA) served as the negative control for microalgal growth and promoting metabolites because it does not have any plant growth-promoting effects; it also served as a positive control in the CO2 experiment because it produces CO2 , as any E. coli . .. Bacteria were then harvested by centrifugation at 3720 × g for 7 min, rinsed twice in 0.85% saline solution, and subsequently transferred to minimal mineral Brunner’s medium (DSMZ medium 457) composed of (in g L−1 ): Na2 HPO4 (2.44), KH2 PO4 (1.52), (NH4 )2 SO4 (0.50), MgSO4 •7H2 O (0.20), CaCl2 •2H2 O (0.05), EDTA (0.50), FeSO4 •7H2 O (0.20), and (in μg·L−1 ): ZnSO4 •7H2 O (0.10), MnCl2 •4H2 O (0.03), H3 BO3 (0.30), CoCl2 •6H2 O (0.20), CuCl2 •2H2 O (0.01), NiCl2 •6H2 O (0.02), Na2 MoO4 •2H2 O (0.03).

    Amplification:

    Article Title: Low expression of PinX1 is associated with malignant behavior in basal-like breast cancer
    Article Snippet: E. coli DH5α (Invitrogen, Carlsbad, CA, USA) was maintained in flasks with LB medium. .. The plasmid pSpCas9(BB)-2A-GFP (PX458) for gene knockout was aquired from Feng Zhang Lab (#48138; Addgene, Cambridge, MA, USA). pCDNA3.1(+) (Invitrogen) and pEGFP-N1 vectors (Clontech Laboratories, Inc., Mountain View, CA, USA) were used for gene amplification in mammalian cells.

    Article Title: Feedback Inhibition of Chorismate Mutase/Prephenate Dehydrogenase (TyrA) of Escherichia coli: Generation and Characterization of Tyrosine-Insensitive Mutants
    Article Snippet: Putative tyrA fbr genes were amplified by PCR using the following primers: tyrA_LIC_fw (5′-GGT ATT GAG GGT CGC ATG GTT GCT GAA TTG ACC GCA TTA C-3′) and tyrA_LIC_rv (5′-AGA GGA GAG TTA GAG CCT TAT TAC TGG CGA TTG TCA TTC GCC-3′). .. After gel purification, the PCR products were subcloned into pET-30 Xa/LIC (Novagen) by using ligation-independent cloning (LIC; Novagen) and transformed E. coli DH5α (Invitrogen).

    Article Title: Aggregating sequences that occur in many proteins constitute weak spots of bacterial proteostasis
    Article Snippet: .. Escherichia coli DH5α (Thermo Fisher Scientific) was used for cloning and plasmid amplification. .. For selection of antibiotic resistance colonies, E . coli carrying plasmids was grown in LB medium supplemented with the relevant antibiotic.

    Positive Control:

    Article Title: Enhanced performance of the microalga Chlorella sorokiniana remotely induced by the plant growth-promoting bacteria Azospirillum brasilense and Bacillus pumilus
    Article Snippet: .. In experiments that measured the potential effect of CO2 , Escherichia coli DH5α (Invitrogen, Carlsbad, CA) served as the negative control for microalgal growth and promoting metabolites because it does not have any plant growth-promoting effects; it also served as a positive control in the CO2 experiment because it produces CO2 , as any E. coli . .. For initial culturing of the microalga, 10 mL axenic C. sorokiniana culture, cultivated in sterile mineral medium (C30), was added to a sterile flask containing 90 mL sterile C30 medium, composed of (in g·L−1 ): KNO3 (25), MgSO4 •7H2 O (10), KH2 PO4 (4), K2 HPO4 (1), FeSO4 •7H2 O (1), and (in μg·L−1 ): H3 BO3 (2.86), MnCl2 •4H2 O (1.81), ZnSO4 •7H2 O (0.11), CuSO4 •5H2 O (0.09), NaMoO4 (0.021), pH 5.25 and incubated at 27 ± 2 °C on a rotary shaker at 120 rpm under 60 μmol photon·m−2 ·s−1 continuous light intensity for 6 days .

    Incubation:

    Article Title: Enhanced performance of the microalga Chlorella sorokiniana remotely induced by the plant growth-promoting bacteria Azospirillum brasilense and Bacillus pumilus
    Article Snippet: In experiments that measured the potential effect of CO2 , Escherichia coli DH5α (Invitrogen, Carlsbad, CA) served as the negative control for microalgal growth and promoting metabolites because it does not have any plant growth-promoting effects; it also served as a positive control in the CO2 experiment because it produces CO2 , as any E. coli . .. For initial culturing of the microalga, 10 mL axenic C. sorokiniana culture, cultivated in sterile mineral medium (C30), was added to a sterile flask containing 90 mL sterile C30 medium, composed of (in g·L−1 ): KNO3 (25), MgSO4 •7H2 O (10), KH2 PO4 (4), K2 HPO4 (1), FeSO4 •7H2 O (1), and (in μg·L−1 ): H3 BO3 (2.86), MnCl2 •4H2 O (1.81), ZnSO4 •7H2 O (0.11), CuSO4 •5H2 O (0.09), NaMoO4 (0.021), pH 5.25 and incubated at 27 ± 2 °C on a rotary shaker at 120 rpm under 60 μmol photon·m−2 ·s−1 continuous light intensity for 6 days .

    Article Title: Characterization of a minimal pKW2124 replicon from Weissella cibaria KLC140 and its application for the construction of the Weissella expression vector pKUCm1
    Article Snippet: .. All Weissella species were incubated anaerobically at 37°C in de Man-Rogosa-Sharpe (MRS) medium (Difco, Detroit, MI, USA) and E. coli DH5α (Invitrogen, Carlsbad, CA, USA) was grown with shaking in Luria-Bertani (LB) medium (Difco) at 37°C. .. Ampicillin sulfate (Sigma, St. Louis, MO, USA) was added to the E. coli growth medium for selection at 50 μg/ml.

    Activity Assay:

    Article Title: Reduced Airway Surface pH Impairs Bacterial Killing in the Porcine Cystic Fibrosis Lung
    Article Snippet: Luminescence antibacterial assay To assess the effect of pH on the antibacterial activity of lysozyme and lactoferrin, we used a luminescence assay of bacterial viability that has been described previously . .. The bacteria used were S. aureus Xen29 (Caliper LifeSciences Bioware™, Hopkinton, MA) and E. coli DH5α (GIBCO BRL, Life Technologies, Grand Island, NY) expressing a luminescence plasmid pCGLS1.

    Cell Culture:

    Article Title: What an Escherichia coli Mutant Can Teach Us About the Antibacterial Effect of Chlorophyllin
    Article Snippet: .. Bacteria Strains and Cell Culture Experiments were performed with E. coli DH5α (Invitrogen, Carlsbad, CA, USA), E. coli NR698 (MC4100 lptD4213 ; kindly provided by M. Grabowicz, Princeton University, NJ, USA) [ ] and B. subtilis 168 (trpC2 ; laboratory stock). ..

    Article Title: Low expression of PinX1 is associated with malignant behavior in basal-like breast cancer
    Article Snippet: It was cultured in Dulbeccos modified Eagles medium (DMEM; Gibco, Carlsbad, CA, USA) containing 10% fetal bovine serum (FBS; Gibco) at 37°C and 5% CO2 . .. E. coli DH5α (Invitrogen, Carlsbad, CA, USA) was maintained in flasks with LB medium.

    Article Title: Chronic Prosthetic Hip Infection Caused by a Small-Colony Variant of Escherichia coli
    Article Snippet: E. coli DH5α was purchased from Gibco-BRL (Eggenstein, Germany). .. Unless otherwise stated, bacteria were cultured on Trypticase soy agar (TSA; Oxoid, Unipath Ltd., Basingstoke, England) supplemented with 7% defibrinated sheep blood (Oxoid, Unipath Ltd.) for 48 h at 37°C under aerobic conditions.

    Expressing:

    Article Title: Reduced Airway Surface pH Impairs Bacterial Killing in the Porcine Cystic Fibrosis Lung
    Article Snippet: .. The bacteria used were S. aureus Xen29 (Caliper LifeSciences Bioware™, Hopkinton, MA) and E. coli DH5α (GIBCO BRL, Life Technologies, Grand Island, NY) expressing a luminescence plasmid pCGLS1. .. E. coli were grown in Luria-Bertani medium containing ampicillin (100 µg/ml, to maintain the plasmid) at 30°C with shaking.

    Article Title: The multi-catalytic compartment of propionyl-CoA synthase sequesters a toxic metabolite
    Article Snippet: E. coli DH5α (Thermo Scientific™) strains were used for cloning and grown in LB medium . .. For protein expression E. coli BL21-AI™ (Invitrogen) or Arctic-Express (DE3) RIL (Agilent Technologies) were grown in TB medium .

    Article Title: Kinetic Properties of Four Plasmid-Mediated AmpC ?-Lactamases
    Article Snippet: .. E. coli DH5α (Gibco-BRL-Life Technologies, Eragny, France) and E. coli BL21(DE3) (Novagen Inc., Madison, WI) were used as recipients for cloning and expression experiments, respectively. .. The plasmid pGEM-T-easy from Promega (Madison, WI) was used to clone the PCR products and pET26b(+) (Novagen, Inc.) was used to produce the different plasmid-encoded class C β-lactamases.

    Modification:

    Article Title: Low expression of PinX1 is associated with malignant behavior in basal-like breast cancer
    Article Snippet: It was cultured in Dulbeccos modified Eagles medium (DMEM; Gibco, Carlsbad, CA, USA) containing 10% fetal bovine serum (FBS; Gibco) at 37°C and 5% CO2 . .. E. coli DH5α (Invitrogen, Carlsbad, CA, USA) was maintained in flasks with LB medium.

    Transformation Assay:

    Article Title: Feedback Inhibition of Chorismate Mutase/Prephenate Dehydrogenase (TyrA) of Escherichia coli: Generation and Characterization of Tyrosine-Insensitive Mutants
    Article Snippet: .. After gel purification, the PCR products were subcloned into pET-30 Xa/LIC (Novagen) by using ligation-independent cloning (LIC; Novagen) and transformed E. coli DH5α (Invitrogen). .. The resulting plasmids, pET30:: tyrA fbr , were analyzed by DNA sequencing with the following primers: T7_prom (5′-TAA TAC GAC TCA CTA TAG GG-3′), T7_term (5′-GCT AGT TAT TGC TCA GCG G-3′), tyrA_291fw (5′-ACT GCG TCC GGT GGT TAT CG-3′), and tyrA_913rv (5′-GGC GAA GAG AGC GCC AGA AG-3′).

    Gel Purification:

    Article Title: Feedback Inhibition of Chorismate Mutase/Prephenate Dehydrogenase (TyrA) of Escherichia coli: Generation and Characterization of Tyrosine-Insensitive Mutants
    Article Snippet: .. After gel purification, the PCR products were subcloned into pET-30 Xa/LIC (Novagen) by using ligation-independent cloning (LIC; Novagen) and transformed E. coli DH5α (Invitrogen). .. The resulting plasmids, pET30:: tyrA fbr , were analyzed by DNA sequencing with the following primers: T7_prom (5′-TAA TAC GAC TCA CTA TAG GG-3′), T7_term (5′-GCT AGT TAT TGC TCA GCG G-3′), tyrA_291fw (5′-ACT GCG TCC GGT GGT TAT CG-3′), and tyrA_913rv (5′-GGC GAA GAG AGC GCC AGA AG-3′).

    Electroporation:

    Article Title: Characterization of a minimal pKW2124 replicon from Weissella cibaria KLC140 and its application for the construction of the Weissella expression vector pKUCm1
    Article Snippet: All Weissella species were incubated anaerobically at 37°C in de Man-Rogosa-Sharpe (MRS) medium (Difco, Detroit, MI, USA) and E. coli DH5α (Invitrogen, Carlsbad, CA, USA) was grown with shaking in Luria-Bertani (LB) medium (Difco) at 37°C. .. Weissella and other electroporation host bacteria were selected using chloramphenicol (USB Corporation, Santa Clara, CA, USA) with the following appropriate concentrations, 6.0 μg/ml for Weissella, Lactobacillus and Bifidobacterium, 5.0 μg/ml for Lactococcus and Leuconostoc , and 3.0 μg/ml for Streptococcus .

    Ligation:

    Article Title: Feedback Inhibition of Chorismate Mutase/Prephenate Dehydrogenase (TyrA) of Escherichia coli: Generation and Characterization of Tyrosine-Insensitive Mutants
    Article Snippet: .. After gel purification, the PCR products were subcloned into pET-30 Xa/LIC (Novagen) by using ligation-independent cloning (LIC; Novagen) and transformed E. coli DH5α (Invitrogen). .. The resulting plasmids, pET30:: tyrA fbr , were analyzed by DNA sequencing with the following primers: T7_prom (5′-TAA TAC GAC TCA CTA TAG GG-3′), T7_term (5′-GCT AGT TAT TGC TCA GCG G-3′), tyrA_291fw (5′-ACT GCG TCC GGT GGT TAT CG-3′), and tyrA_913rv (5′-GGC GAA GAG AGC GCC AGA AG-3′).

    Infection:

    Article Title: Chronic Prosthetic Hip Infection Caused by a Small-Colony Variant of Escherichia coli
    Article Snippet: E. coli Z-2376 was obtained from two different specimens of the scar area of a patient with a prosthetic hip infection. .. E. coli DH5α was purchased from Gibco-BRL (Eggenstein, Germany).

    Article Title: Use of Chimeric Type IV Secretion Systems to Define Contributions of Outer Membrane Subassemblies for Contact-Dependent Translocation
    Article Snippet: E. coli DH5α (GIBCO-BRL) was used for plasmid constructions and the type VI secretion system (T6SS) killing assay. .. E. coli MG1655 ( E. coli Genetic Stock Center) served as donors in the conjugation assays and for the phage infection assays.

    DNA Sequencing:

    Article Title: Feedback Inhibition of Chorismate Mutase/Prephenate Dehydrogenase (TyrA) of Escherichia coli: Generation and Characterization of Tyrosine-Insensitive Mutants
    Article Snippet: Paragraph title: Subcloning of tyrA and DNA sequencing. ... After gel purification, the PCR products were subcloned into pET-30 Xa/LIC (Novagen) by using ligation-independent cloning (LIC; Novagen) and transformed E. coli DH5α (Invitrogen).

    Polymerase Chain Reaction:

    Article Title: Characterization of a minimal pKW2124 replicon from Weissella cibaria KLC140 and its application for the construction of the Weissella expression vector pKUCm1
    Article Snippet: Paragraph title: BACTERIAL STRAINS, PLASMIDS, PCR PRIMERS, AND GROWTH CONDITIONS ... All Weissella species were incubated anaerobically at 37°C in de Man-Rogosa-Sharpe (MRS) medium (Difco, Detroit, MI, USA) and E. coli DH5α (Invitrogen, Carlsbad, CA, USA) was grown with shaking in Luria-Bertani (LB) medium (Difco) at 37°C.

    Article Title: Feedback Inhibition of Chorismate Mutase/Prephenate Dehydrogenase (TyrA) of Escherichia coli: Generation and Characterization of Tyrosine-Insensitive Mutants
    Article Snippet: .. After gel purification, the PCR products were subcloned into pET-30 Xa/LIC (Novagen) by using ligation-independent cloning (LIC; Novagen) and transformed E. coli DH5α (Invitrogen). .. The resulting plasmids, pET30:: tyrA fbr , were analyzed by DNA sequencing with the following primers: T7_prom (5′-TAA TAC GAC TCA CTA TAG GG-3′), T7_term (5′-GCT AGT TAT TGC TCA GCG G-3′), tyrA_291fw (5′-ACT GCG TCC GGT GGT TAT CG-3′), and tyrA_913rv (5′-GGC GAA GAG AGC GCC AGA AG-3′).

    Article Title: Kinetic Properties of Four Plasmid-Mediated AmpC ?-Lactamases
    Article Snippet: E. coli DH5α (Gibco-BRL-Life Technologies, Eragny, France) and E. coli BL21(DE3) (Novagen Inc., Madison, WI) were used as recipients for cloning and expression experiments, respectively. .. The plasmid pGEM-T-easy from Promega (Madison, WI) was used to clone the PCR products and pET26b(+) (Novagen, Inc.) was used to produce the different plasmid-encoded class C β-lactamases.

    Gene Knockout:

    Article Title: Low expression of PinX1 is associated with malignant behavior in basal-like breast cancer
    Article Snippet: E. coli DH5α (Invitrogen, Carlsbad, CA, USA) was maintained in flasks with LB medium. .. The plasmid pSpCas9(BB)-2A-GFP (PX458) for gene knockout was aquired from Feng Zhang Lab (#48138; Addgene, Cambridge, MA, USA). pCDNA3.1(+) (Invitrogen) and pEGFP-N1 vectors (Clontech Laboratories, Inc., Mountain View, CA, USA) were used for gene amplification in mammalian cells.

    Conjugation Assay:

    Article Title: Use of Chimeric Type IV Secretion Systems to Define Contributions of Outer Membrane Subassemblies for Contact-Dependent Translocation
    Article Snippet: E. coli DH5α (GIBCO-BRL) was used for plasmid constructions and the type VI secretion system (T6SS) killing assay. .. E. coli MG1655 ( E. coli Genetic Stock Center) served as donors in the conjugation assays and for the phage infection assays.

    Multiple Displacement Amplification:

    Article Title: Low expression of PinX1 is associated with malignant behavior in basal-like breast cancer
    Article Snippet: Cell cultures and plasmids Breast cancer cell line MDA-MB-231 was obtained from laboratory preservation. .. E. coli DH5α (Invitrogen, Carlsbad, CA, USA) was maintained in flasks with LB medium.

    Isolation:

    Article Title: Feedback Inhibition of Chorismate Mutase/Prephenate Dehydrogenase (TyrA) of Escherichia coli: Generation and Characterization of Tyrosine-Insensitive Mutants
    Article Snippet: Paragraph title: Isolation, manipulation and transfer of DNA. ... Plasmid DNA was transferred to chemically competent cells of E. coli DH5α (Invitrogen) and E. coli BL21(DE3) (Novagen), respectively.

    Subcloning:

    Article Title: Feedback Inhibition of Chorismate Mutase/Prephenate Dehydrogenase (TyrA) of Escherichia coli: Generation and Characterization of Tyrosine-Insensitive Mutants
    Article Snippet: Paragraph title: Subcloning of tyrA and DNA sequencing. ... After gel purification, the PCR products were subcloned into pET-30 Xa/LIC (Novagen) by using ligation-independent cloning (LIC; Novagen) and transformed E. coli DH5α (Invitrogen).

    Microscopy:

    Article Title: Chronic Prosthetic Hip Infection Caused by a Small-Colony Variant of Escherichia coli
    Article Snippet: E. coli DH5α was purchased from Gibco-BRL (Eggenstein, Germany). .. The sizes of the colonies were determined by plate microscopy.

    Purification:

    Article Title: Feedback Inhibition of Chorismate Mutase/Prephenate Dehydrogenase (TyrA) of Escherichia coli: Generation and Characterization of Tyrosine-Insensitive Mutants
    Article Snippet: Agarose gel purification of DNA fragments was done with the Geneclean spin kit (Q-Biogene). .. Plasmid DNA was transferred to chemically competent cells of E. coli DH5α (Invitrogen) and E. coli BL21(DE3) (Novagen), respectively.

    Selection:

    Article Title: The multi-catalytic compartment of propionyl-CoA synthase sequesters a toxic metabolite
    Article Snippet: E. coli DH5α (Thermo Scientific™) strains were used for cloning and grown in LB medium . .. Antibiotics for selection purposes were used accordingly: 100 µg/ml ampicillin, 15 µg/ml gentamycin.

    Article Title: Characterization of a minimal pKW2124 replicon from Weissella cibaria KLC140 and its application for the construction of the Weissella expression vector pKUCm1
    Article Snippet: All Weissella species were incubated anaerobically at 37°C in de Man-Rogosa-Sharpe (MRS) medium (Difco, Detroit, MI, USA) and E. coli DH5α (Invitrogen, Carlsbad, CA, USA) was grown with shaking in Luria-Bertani (LB) medium (Difco) at 37°C. .. Ampicillin sulfate (Sigma, St. Louis, MO, USA) was added to the E. coli growth medium for selection at 50 μg/ml.

    Article Title: Aggregating sequences that occur in many proteins constitute weak spots of bacterial proteostasis
    Article Snippet: Escherichia coli DH5α (Thermo Fisher Scientific) was used for cloning and plasmid amplification. .. For selection of antibiotic resistance colonies, E . coli carrying plasmids was grown in LB medium supplemented with the relevant antibiotic.

    Activated Clotting Time Assay:

    Article Title: Kinetic Properties of Four Plasmid-Mediated AmpC ?-Lactamases
    Article Snippet: The strains Escherichia coli SNO3 pMG232 (MIR-1 producer), E. coli XL1 pMG261 (ACT-1 producer), and E. coli XL1 pMG262 (CMY-2 producer) were gifts of G. A. Jacoby (Section of Infectious Disease, Lahey Clinic, Burlington, MA). .. E. coli DH5α (Gibco-BRL-Life Technologies, Eragny, France) and E. coli BL21(DE3) (Novagen Inc., Madison, WI) were used as recipients for cloning and expression experiments, respectively.

    Plasmid Preparation:

    Article Title: Reduced Airway Surface pH Impairs Bacterial Killing in the Porcine Cystic Fibrosis Lung
    Article Snippet: .. The bacteria used were S. aureus Xen29 (Caliper LifeSciences Bioware™, Hopkinton, MA) and E. coli DH5α (GIBCO BRL, Life Technologies, Grand Island, NY) expressing a luminescence plasmid pCGLS1. .. E. coli were grown in Luria-Bertani medium containing ampicillin (100 µg/ml, to maintain the plasmid) at 30°C with shaking.

    Article Title: Feedback Inhibition of Chorismate Mutase/Prephenate Dehydrogenase (TyrA) of Escherichia coli: Generation and Characterization of Tyrosine-Insensitive Mutants
    Article Snippet: .. Plasmid DNA was transferred to chemically competent cells of E. coli DH5α (Invitrogen) and E. coli BL21(DE3) (Novagen), respectively. .. The wild-type tyrA gene was amplified by PCR from chromosomal DNA of E. coli K-12 using the following primers: tyrA_fw_KpnI (5′-CCG GTA CCA TGG TTG CTG AAT TGA CCG CAT TAC −3′) and tyrA_rev_MluI (5′-CCA CGC GTT TAT TAC TGG CGA TTG TCA TTC GCC-3′).

    Article Title: Low expression of PinX1 is associated with malignant behavior in basal-like breast cancer
    Article Snippet: E. coli DH5α (Invitrogen, Carlsbad, CA, USA) was maintained in flasks with LB medium. .. The plasmid pSpCas9(BB)-2A-GFP (PX458) for gene knockout was aquired from Feng Zhang Lab (#48138; Addgene, Cambridge, MA, USA). pCDNA3.1(+) (Invitrogen) and pEGFP-N1 vectors (Clontech Laboratories, Inc., Mountain View, CA, USA) were used for gene amplification in mammalian cells.

    Article Title: Transfer of Herb-Resistance Plasmid From Escherichia coli to Staphylococcus aureus Residing in the Human Urinary Tract
    Article Snippet: .. Plasmid Characterization Total plasmid DNA was prepared (Plasmid Extraction Kit, Beijing TIANGEN) and transferred into E. coli DH5α (Invitrogen, Carlsbad, CA, USA). ..

    Article Title: Kinetic Properties of Four Plasmid-Mediated AmpC ?-Lactamases
    Article Snippet: E. coli DH5α (Gibco-BRL-Life Technologies, Eragny, France) and E. coli BL21(DE3) (Novagen Inc., Madison, WI) were used as recipients for cloning and expression experiments, respectively. .. The plasmid pGEM-T-easy from Promega (Madison, WI) was used to clone the PCR products and pET26b(+) (Novagen, Inc.) was used to produce the different plasmid-encoded class C β-lactamases.

    Article Title: Use of Chimeric Type IV Secretion Systems to Define Contributions of Outer Membrane Subassemblies for Contact-Dependent Translocation
    Article Snippet: .. E. coli DH5α (GIBCO-BRL) was used for plasmid constructions and the type VI secretion system (T6SS) killing assay. .. E. coli MG1655 ( E. coli Genetic Stock Center) served as donors in the conjugation assays and for the phage infection assays.

    Article Title: Aggregating sequences that occur in many proteins constitute weak spots of bacterial proteostasis
    Article Snippet: .. Escherichia coli DH5α (Thermo Fisher Scientific) was used for cloning and plasmid amplification. .. For selection of antibiotic resistance colonies, E . coli carrying plasmids was grown in LB medium supplemented with the relevant antibiotic.

    Software:

    Article Title: The multi-catalytic compartment of propionyl-CoA synthase sequesters a toxic metabolite
    Article Snippet: E. coli DH5α (Thermo Scientific™) strains were used for cloning and grown in LB medium . .. All in silico cloning was performed with Clone Manager 9 (Scientific & Educational Software).

    Negative Control:

    Article Title: Enhanced performance of the microalga Chlorella sorokiniana remotely induced by the plant growth-promoting bacteria Azospirillum brasilense and Bacillus pumilus
    Article Snippet: .. In experiments that measured the potential effect of CO2 , Escherichia coli DH5α (Invitrogen, Carlsbad, CA) served as the negative control for microalgal growth and promoting metabolites because it does not have any plant growth-promoting effects; it also served as a positive control in the CO2 experiment because it produces CO2 , as any E. coli . .. For initial culturing of the microalga, 10 mL axenic C. sorokiniana culture, cultivated in sterile mineral medium (C30), was added to a sterile flask containing 90 mL sterile C30 medium, composed of (in g·L−1 ): KNO3 (25), MgSO4 •7H2 O (10), KH2 PO4 (4), K2 HPO4 (1), FeSO4 •7H2 O (1), and (in μg·L−1 ): H3 BO3 (2.86), MnCl2 •4H2 O (1.81), ZnSO4 •7H2 O (0.11), CuSO4 •5H2 O (0.09), NaMoO4 (0.021), pH 5.25 and incubated at 27 ± 2 °C on a rotary shaker at 120 rpm under 60 μmol photon·m−2 ·s−1 continuous light intensity for 6 days .

    Positron Emission Tomography:

    Article Title: Feedback Inhibition of Chorismate Mutase/Prephenate Dehydrogenase (TyrA) of Escherichia coli: Generation and Characterization of Tyrosine-Insensitive Mutants
    Article Snippet: .. After gel purification, the PCR products were subcloned into pET-30 Xa/LIC (Novagen) by using ligation-independent cloning (LIC; Novagen) and transformed E. coli DH5α (Invitrogen). .. The resulting plasmids, pET30:: tyrA fbr , were analyzed by DNA sequencing with the following primers: T7_prom (5′-TAA TAC GAC TCA CTA TAG GG-3′), T7_term (5′-GCT AGT TAT TGC TCA GCG G-3′), tyrA_291fw (5′-ACT GCG TCC GGT GGT TAT CG-3′), and tyrA_913rv (5′-GGC GAA GAG AGC GCC AGA AG-3′).

    Agarose Gel Electrophoresis:

    Article Title: Feedback Inhibition of Chorismate Mutase/Prephenate Dehydrogenase (TyrA) of Escherichia coli: Generation and Characterization of Tyrosine-Insensitive Mutants
    Article Snippet: Agarose gel purification of DNA fragments was done with the Geneclean spin kit (Q-Biogene). .. Plasmid DNA was transferred to chemically competent cells of E. coli DH5α (Invitrogen) and E. coli BL21(DE3) (Novagen), respectively.

    Article Title: Transfer of Herb-Resistance Plasmid From Escherichia coli to Staphylococcus aureus Residing in the Human Urinary Tract
    Article Snippet: Plasmid Characterization Total plasmid DNA was prepared (Plasmid Extraction Kit, Beijing TIANGEN) and transferred into E. coli DH5α (Invitrogen, Carlsbad, CA, USA). .. Plasmid DNA was separated on a 0.7% agarose gel, stained with ethidium bromide, and visualized under UV-light.

    Preserving:

    Article Title: Low expression of PinX1 is associated with malignant behavior in basal-like breast cancer
    Article Snippet: Cell cultures and plasmids Breast cancer cell line MDA-MB-231 was obtained from laboratory preservation. .. E. coli DH5α (Invitrogen, Carlsbad, CA, USA) was maintained in flasks with LB medium.

    Nuclear Magnetic Resonance:

    Article Title: RNA polymerase-induced remodelling of NusA produces a pause enhancement complex
    Article Snippet: DNA manipulation All cloning was carried out using E. coli DH5α (Gibco BRL; Supplementary Table S1). .. A fragment of B. subtilis rpoB containing the β-flap (residues 784–923; corresponding to residues 830–1058 of the E. coli β subunit) was inserted into pETMCSIII to produce pNG812 (Supplementary Table S1) for overproduction of protein used in NMR studies.

    Concentration Assay:

    Article Title: What an Escherichia coli Mutant Can Teach Us About the Antibacterial Effect of Chlorophyllin
    Article Snippet: Bacteria Strains and Cell Culture Experiments were performed with E. coli DH5α (Invitrogen, Carlsbad, CA, USA), E. coli NR698 (MC4100 lptD4213 ; kindly provided by M. Grabowicz, Princeton University, NJ, USA) [ ] and B. subtilis 168 (trpC2 ; laboratory stock). .. Prior to the experiments, cell concentration was determined optically at 590 nm and set to OD590 = 0.1 before cells were diluted in LB as required.

    DNA Purification:

    Article Title: Feedback Inhibition of Chorismate Mutase/Prephenate Dehydrogenase (TyrA) of Escherichia coli: Generation and Characterization of Tyrosine-Insensitive Mutants
    Article Snippet: Chromosomal DNA from E. coli K-12 was prepared by using the Wizard genomic DNA purification kit (Promega). .. Plasmid DNA was transferred to chemically competent cells of E. coli DH5α (Invitrogen) and E. coli BL21(DE3) (Novagen), respectively.

    Luminescence Assay:

    Article Title: Reduced Airway Surface pH Impairs Bacterial Killing in the Porcine Cystic Fibrosis Lung
    Article Snippet: Luminescence antibacterial assay To assess the effect of pH on the antibacterial activity of lysozyme and lactoferrin, we used a luminescence assay of bacterial viability that has been described previously . .. The bacteria used were S. aureus Xen29 (Caliper LifeSciences Bioware™, Hopkinton, MA) and E. coli DH5α (GIBCO BRL, Life Technologies, Grand Island, NY) expressing a luminescence plasmid pCGLS1.

    Staining:

    Article Title: Transfer of Herb-Resistance Plasmid From Escherichia coli to Staphylococcus aureus Residing in the Human Urinary Tract
    Article Snippet: Plasmid Characterization Total plasmid DNA was prepared (Plasmid Extraction Kit, Beijing TIANGEN) and transferred into E. coli DH5α (Invitrogen, Carlsbad, CA, USA). .. Plasmid DNA was separated on a 0.7% agarose gel, stained with ethidium bromide, and visualized under UV-light.

    In Silico:

    Article Title: The multi-catalytic compartment of propionyl-CoA synthase sequesters a toxic metabolite
    Article Snippet: E. coli DH5α (Thermo Scientific™) strains were used for cloning and grown in LB medium . .. All in silico cloning was performed with Clone Manager 9 (Scientific & Educational Software).

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