e coli novablue  (Millipore)


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    Structured Review

    Millipore e coli novablue
    E Coli Novablue, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/e coli novablue/product/Millipore
    Average 99 stars, based on 11 article reviews
    Price from $9.99 to $1999.99
    e coli novablue - by Bioz Stars, 2019-12
    99/100 stars

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    Related Articles

    Clone Assay:

    Article Title: Aspergillus oryzae-based cell factory for direct kojic acid production from cellulose
    Article Snippet: The washed mycelium was recultivated in 500 mL Erlenmeyer flasks with baffles containing 100 mL CY medium at 30°C that were shaken at 150 rpm using an orbital shaker. .. E. coli NovaBlue (Novagen, Inc., Madison, WI, USA) was used as the cloning host for recombinant DNA manipulations. .. The bacterium was grown in Luria–Bertani medium (1% tryptone, 0.5% yeast extract, and 0.5% NaCl) containing 0.1 mg/mL of ampicillin.

    Article Title: Treponema denticola PrcB Is Required for Expression and Activity of the PrcA-PrtP (Dentilisin) Complex
    Article Snippet: Cultures were examined by phase-contrast microscopy for purity and typical strain morphology before use. .. Escherichia coli NovaBlue (Novagen, Inc., Madison, WI) and JM109 ( ) were used as hosts for cloning. .. E. coli was grown in LB agar or broth medium with ampicillin (50 μg ml−1 ), kanamycin (30 μg ml−1 ), and Em (200 μg ml−1 ), as appropriate.

    Article Title: Characterization and Recombinant Expression of Terebrid Venom Peptide from Terebra guttata
    Article Snippet: The PCR amplified insert was purified with SpinPrep Gel DNA Kit (Novagen, Darmstadt, Germany) and cloned via LIC into vector pET-32 Xa/LIC (EMD Millipore, Darmstadt, Germany). .. The pET-32 Xa/LIC:Tgu6.1 plasmid construct was transformed into E. coli NovaBlue obtained from Novagen.

    Article Title: Phospholipase D from Loxosceleslaeta Spider Venom Induces IL-6, IL-8, CXCL1/GRO-α, and CCL2/MCP-1 Production in Human Skin Fibroblasts and Stimulates Monocytes Migration
    Article Snippet: Thermal cycling conditions were: 95 °C for 3 min, followed by 32 cycles of 95 °C for 30 s, 58 °C for 30 s, and 72 °C for 1 min, and followed by a final incubation at 72 °C for 5 min. Amplification products were separated in 1.5% agarose gel in TAE buffer and stained with EthBr, then gel images were captured in a DNR Bio-Imaging System MF-ChemiBIS 2.0. (Mahale HaHamisha, Jerusalem, Israel) using Gel-Capture software. .. In addition, each gene was cloned into a cloning vector pCR2.1® -TOPO® using the TOPO TA Cloning® kit (Invitrogen) and transformed into E. coli Novablue (EMD Chemicals-Novagen Brand., Madison, WI, USA) Then, cloned gene plasmids were purified and used to prepare a standard curve for quantitative RT-PCR (RT-qPCR) in real time. .. The real-time efficiencies were determined for each gene from standard curves generated from serial dilutions of plasmids sample.

    Article Title: Proteomic Analysis of and Immune Responses to Ehrlichia chaffeensis Lipoproteins
    Article Snippet: PCR products were cloned into pCR-Blunt II-topo (Invitrogen) according to the manufacturer's instructions. .. The recombinant plasmids were amplified in E. coli NovaBlue (Novagen).

    Article Title: Display of both N- and C-terminal target fusion proteins on the Aspergillus oryzae cell surface using a chitin-binding module
    Article Snippet: Using CBM as an anchor protein, we tested the displaying possibilities of green fluorescent protein (GFP) and triacylglycerol lipase from A. oryzae (tglA). .. Escherichia coli NovaBlue (Novagen, Inc., Madison, WI) was used as the cloning host for recombinant DNA manipulations. .. The bacterium was grown in Luria–Bertani medium (1% tryptone, 0.5% yeast extract, and 0.5% NaCl) containing 0.1 mg/ml of ampicillin.

    Article Title: Equine Arteritis Virus Uses Equine CXCL16 as an Entry Receptor
    Article Snippet: Paragraph title: Cloning and expression of EqCXCL16 in Escherichia coli . ... The resultant plasmid construct (p15-16A) was used to transform E. coli NovaBlue (EMD Millipore Novagen, Temecula, CA) using a TransformAid kit (Thermo Scientific, Rockford, IL).

    Article Title: Diversity of hydrolases from hydrothermal vent sediments of the Levante Bay, Vulcano Island (Aeolian archipelago) identified by activity-based metagenomics and biochemical characterization of new esterases and an arabinopyranosidase
    Article Snippet: Paragraph title: Cloning and expression of selected genes in E. coli ... The purified PCR products were then purified, treated with an endonuclease, annealed to His-tag harbouring the pET-46c Ek/LIC vector, and transformed into E. coli NovaBlue according to the protocol of the manufacturer (Novagen, Darmstadt, Germany).

    Article Title: MprAB Regulates the espA Operon in Mycobacterium tuberculosis and Modulates ESX-1 Function and Host Cytokine Response
    Article Snippet: Strains were grown in Middlebrook 7H9 broth containing 0.05% Tween 80 and 10% oleic acid-albumin-dextrose-catalase (OADC) at 37°C with shaking under atmospheric CO2 . .. Escherichia coli NovaBlue and Rosetta(DE3)pLysS (Novagen) were used for general cloning and protein expression, respectively. .. Briefly, M. tuberculosis strains were grown to mid-exponential phase and total RNA was extracted by bead beating, followed by further purification and DNase treatment.

    Article Title: The ?-propeller gene Rv1057 of Mycobacterium tuberculosis has a complex promoter directly regulated by both the MprAB and TrcRS two-component systems
    Article Snippet: M. tuberculosis strain Rv-D981 is an mprAB deletion mutant derived from the laboratory strain H37Rv, and Rv-D981Com is the mprAB -complemented strain generated from Rv-D981 . .. Escherichia coli Novablue and Rosetta(DE3)pLysS (Novagen) were used for general cloning purposes and protein expression, respectively. .. Rv1057 promoter fusions were constructed in the integrating vector pSM128 by inserting sections of the Rv1056-Rv1057 intergenic region upstream of lacZ .

    Centrifugation:

    Article Title: Equine Arteritis Virus Uses Equine CXCL16 as an Entry Receptor
    Article Snippet: The resultant plasmid construct (p15-16A) was used to transform E. coli NovaBlue (EMD Millipore Novagen, Temecula, CA) using a TransformAid kit (Thermo Scientific, Rockford, IL). .. The resultant plasmid construct (p15-16A) was used to transform E. coli NovaBlue (EMD Millipore Novagen, Temecula, CA) using a TransformAid kit (Thermo Scientific, Rockford, IL).

    Article Title: Diversity of hydrolases from hydrothermal vent sediments of the Levante Bay, Vulcano Island (Aeolian archipelago) identified by activity-based metagenomics and biochemical characterization of new esterases and an arabinopyranosidase
    Article Snippet: The purified PCR products were then purified, treated with an endonuclease, annealed to His-tag harbouring the pET-46c Ek/LIC vector, and transformed into E. coli NovaBlue according to the protocol of the manufacturer (Novagen, Darmstadt, Germany). .. The cultures were then transferred to an incubator at 18 °C and induced by adding isopropyl-β-d -galactopyranoside (IPTG) at a final concentration of 0.5 mM.

    Amplification:

    Article Title: Penicillin-Binding Protein of Ehrlichia chaffeensis: Cytokine Induction Through MyD88-Dependent Pathway
    Article Snippet: The amplified fragments were digested with restriction enzymes and ligated into restriction enzyme–digested pET-33b(+) (Novagen). .. Escherichia coli Novablue (Novagen) was transformed and the plasmids were extracted.

    Article Title: Characterization and Recombinant Expression of Terebrid Venom Peptide from Terebra guttata
    Article Snippet: The PCR amplified insert was purified with SpinPrep Gel DNA Kit (Novagen, Darmstadt, Germany) and cloned via LIC into vector pET-32 Xa/LIC (EMD Millipore, Darmstadt, Germany). .. The pET-32 Xa/LIC:Tgu6.1 plasmid construct was transformed into E. coli NovaBlue obtained from Novagen.

    Article Title: Phospholipase D from Loxosceleslaeta Spider Venom Induces IL-6, IL-8, CXCL1/GRO-α, and CCL2/MCP-1 Production in Human Skin Fibroblasts and Stimulates Monocytes Migration
    Article Snippet: Thermal cycling conditions were: 95 °C for 3 min, followed by 32 cycles of 95 °C for 30 s, 58 °C for 30 s, and 72 °C for 1 min, and followed by a final incubation at 72 °C for 5 min. Amplification products were separated in 1.5% agarose gel in TAE buffer and stained with EthBr, then gel images were captured in a DNR Bio-Imaging System MF-ChemiBIS 2.0. (Mahale HaHamisha, Jerusalem, Israel) using Gel-Capture software. .. In addition, each gene was cloned into a cloning vector pCR2.1® -TOPO® using the TOPO TA Cloning® kit (Invitrogen) and transformed into E. coli Novablue (EMD Chemicals-Novagen Brand., Madison, WI, USA) Then, cloned gene plasmids were purified and used to prepare a standard curve for quantitative RT-PCR (RT-qPCR) in real time.

    Article Title: Integrating artificial with natural cells to translate chemical messages that direct E. coli behaviour
    Article Snippet: Data are reported as averages with standard error, or representative runs are shown. .. Plasmids were amplified in E. coli Novablue (Novagen) and purified with Wizard Plus SV Minipreps DNA Purification System (Promega). .. Plasmid DNA was phenol–chloroform extracted, ethanol precipitated and resuspended in deionized and diethyl pyrocarbonate-treated water.

    Article Title: Proteomic Analysis of and Immune Responses to Ehrlichia chaffeensis Lipoproteins
    Article Snippet: Inserts excised from pCR-Blunt II were directionally subcloned into E. coli expression vector pET29a(+) (Novagen, San Diego, CA) digested with NdeI and XhoI. .. The recombinant plasmids were amplified in E. coli NovaBlue (Novagen). .. Recombinant lipoproteins were expressed in E. coli BL21(DE3).

    Article Title: Equine Arteritis Virus Uses Equine CXCL16 as an Entry Receptor
    Article Snippet: The amplified EqCXCL16 fragment (amino acids [aa] 17 to 247) was run on a 1% agarose gel (E-Gel EX; Life Technologies, Grand Island, NY) and purified with a commercial kit (Zymoclean gel recovery kit; Zymo Research, Irvine, CA). .. The resultant plasmid construct (p15-16A) was used to transform E. coli NovaBlue (EMD Millipore Novagen, Temecula, CA) using a TransformAid kit (Thermo Scientific, Rockford, IL).

    Filtration:

    Article Title: Aspergillus oryzae-based cell factory for direct kojic acid production from cellulose
    Article Snippet: The mycelium in culture medium was separated from the culture medium by filtration using Miracloth (Calbiochem, Darmstadt, Germany) and was washed by distilled water. .. E. coli NovaBlue (Novagen, Inc., Madison, WI, USA) was used as the cloning host for recombinant DNA manipulations.

    Recovery:

    Article Title: Equine Arteritis Virus Uses Equine CXCL16 as an Entry Receptor
    Article Snippet: The amplified EqCXCL16 fragment (amino acids [aa] 17 to 247) was run on a 1% agarose gel (E-Gel EX; Life Technologies, Grand Island, NY) and purified with a commercial kit (Zymoclean gel recovery kit; Zymo Research, Irvine, CA). .. The resultant plasmid construct (p15-16A) was used to transform E. coli NovaBlue (EMD Millipore Novagen, Temecula, CA) using a TransformAid kit (Thermo Scientific, Rockford, IL).

    DNA Ligation:

    Article Title: Equine Arteritis Virus Uses Equine CXCL16 as an Entry Receptor
    Article Snippet: The purified EqCXCL16 fragment was digested with the XhoI and BamHI restriction enzymes and ligated (Rapid DNA ligation kit; Thermo Scientific, Rockford, IL) into the bacterial expression vector pET15b (EMD Millipore Novagen, Temecula, CA) following digestion with the same restriction enzymes. .. The resultant plasmid construct (p15-16A) was used to transform E. coli NovaBlue (EMD Millipore Novagen, Temecula, CA) using a TransformAid kit (Thermo Scientific, Rockford, IL).

    Polymerase Chain Reaction:

    Article Title: Penicillin-Binding Protein of Ehrlichia chaffeensis: Cytokine Induction Through MyD88-Dependent Pathway
    Article Snippet: The DNA fragments encoding the full-length Ech_1067 without signal peptide (1–30 aa) and the full-length diaminopimelate aminotransferase (DPA, Ech_0886) were amplified by polymerase chain reaction (PCR) using E. chaffeensis chromosomal DNA as a template. .. Escherichia coli Novablue (Novagen) was transformed and the plasmids were extracted.

    Article Title: Treponema denticola PrcB Is Required for Expression and Activity of the PrcA-PrtP (Dentilisin) Complex
    Article Snippet: Escherichia coli NovaBlue (Novagen, Inc., Madison, WI) and JM109 ( ) were used as hosts for cloning. .. E. coli was grown in LB agar or broth medium with ampicillin (50 μg ml−1 ), kanamycin (30 μg ml−1 ), and Em (200 μg ml−1 ), as appropriate.

    Article Title: Characterization and Recombinant Expression of Terebrid Venom Peptide from Terebra guttata
    Article Snippet: The PCR amplified insert was purified with SpinPrep Gel DNA Kit (Novagen, Darmstadt, Germany) and cloned via LIC into vector pET-32 Xa/LIC (EMD Millipore, Darmstadt, Germany). .. The pET-32 Xa/LIC:Tgu6.1 plasmid construct was transformed into E. coli NovaBlue obtained from Novagen.

    Article Title: Phospholipase D from Loxosceleslaeta Spider Venom Induces IL-6, IL-8, CXCL1/GRO-α, and CCL2/MCP-1 Production in Human Skin Fibroblasts and Stimulates Monocytes Migration
    Article Snippet: In addition, each gene was cloned into a cloning vector pCR2.1® -TOPO® using the TOPO TA Cloning® kit (Invitrogen) and transformed into E. coli Novablue (EMD Chemicals-Novagen Brand., Madison, WI, USA) Then, cloned gene plasmids were purified and used to prepare a standard curve for quantitative RT-PCR (RT-qPCR) in real time. .. Then, RT-qPCR was then performed using a thermal cycler StepOnePlus™ Real-Time PCR System (Applied Biosystems., Foster City, CA, USA).

    Article Title: Integrating artificial with natural cells to translate chemical messages that direct E. coli behaviour
    Article Snippet: Plasmids were amplified in E. coli Novablue (Novagen) and purified with Wizard Plus SV Minipreps DNA Purification System (Promega). .. Plasmid DNA was phenol–chloroform extracted, ethanol precipitated and resuspended in deionized and diethyl pyrocarbonate-treated water.

    Article Title: Proteomic Analysis of and Immune Responses to Ehrlichia chaffeensis Lipoproteins
    Article Snippet: Inserts excised from pCR-Blunt II were directionally subcloned into E. coli expression vector pET29a(+) (Novagen, San Diego, CA) digested with NdeI and XhoI. .. The recombinant plasmids were amplified in E. coli NovaBlue (Novagen).

    Article Title: Equine Arteritis Virus Uses Equine CXCL16 as an Entry Receptor
    Article Snippet: The cDNA amplification was carried out according to a standard laboratory PCR protocol using forward primer cx16-15F (5′-GCG CTCGAG GCGTTGCTGACTCTGCAAGG-3′) and reverse primer cx16-15R (5′-GC GGATCC GCACTGCCACTGTAACTGAT-3′) (IDT, Coralville, IA). .. The resultant plasmid construct (p15-16A) was used to transform E. coli NovaBlue (EMD Millipore Novagen, Temecula, CA) using a TransformAid kit (Thermo Scientific, Rockford, IL).

    Article Title: Diversity of hydrolases from hydrothermal vent sediments of the Levante Bay, Vulcano Island (Aeolian archipelago) identified by activity-based metagenomics and biochemical characterization of new esterases and an arabinopyranosidase
    Article Snippet: The reactions were done in a Techne TC-5000 cycler (CA, USA) using the following programme: 1 cycle of 95 °C/3 min following 35 cycles of 95 °C for 30s, 55 °C for 30s, and 72 °C for 1 min per 1000 nucleotides, followed by one extension at 72 °C for 5 min. .. The purified PCR products were then purified, treated with an endonuclease, annealed to His-tag harbouring the pET-46c Ek/LIC vector, and transformed into E. coli NovaBlue according to the protocol of the manufacturer (Novagen, Darmstadt, Germany). .. The DNA inserts in the resulting plasmids were verified by sequencing services of Macrogen Ltd. (Amsterdam, The Netherlands).

    TA Cloning:

    Article Title: Phospholipase D from Loxosceleslaeta Spider Venom Induces IL-6, IL-8, CXCL1/GRO-α, and CCL2/MCP-1 Production in Human Skin Fibroblasts and Stimulates Monocytes Migration
    Article Snippet: Thermal cycling conditions were: 95 °C for 3 min, followed by 32 cycles of 95 °C for 30 s, 58 °C for 30 s, and 72 °C for 1 min, and followed by a final incubation at 72 °C for 5 min. Amplification products were separated in 1.5% agarose gel in TAE buffer and stained with EthBr, then gel images were captured in a DNR Bio-Imaging System MF-ChemiBIS 2.0. (Mahale HaHamisha, Jerusalem, Israel) using Gel-Capture software. .. In addition, each gene was cloned into a cloning vector pCR2.1® -TOPO® using the TOPO TA Cloning® kit (Invitrogen) and transformed into E. coli Novablue (EMD Chemicals-Novagen Brand., Madison, WI, USA) Then, cloned gene plasmids were purified and used to prepare a standard curve for quantitative RT-PCR (RT-qPCR) in real time. .. The real-time efficiencies were determined for each gene from standard curves generated from serial dilutions of plasmids sample.

    Construct:

    Article Title: Enzymatic improvement of mitochondrial thiol oxidase Erv1 for oxidized glutathione fermentation by Saccharomyces cerevisiae
    Article Snippet: Saccharomyces cerevisiae GCIΔGLR1 , GSH1 /GSH2 cocktail δ-integrated and GLR1 deleted YPH499 (ABC1193/NBRC 10505) strain was previously constructed [ ] and used for glutathione production in this study. .. Escherichia coli NovaBlue (Novagen, Darmstadt, Germany) strain was used for DNA manipulation.

    Article Title: Treponema denticola PrcB Is Required for Expression and Activity of the PrcA-PrtP (Dentilisin) Complex
    Article Snippet: Escherichia coli NovaBlue (Novagen, Inc., Madison, WI) and JM109 ( ) were used as hosts for cloning. .. E. coli was grown in LB agar or broth medium with ampicillin (50 μg ml−1 ), kanamycin (30 μg ml−1 ), and Em (200 μg ml−1 ), as appropriate.

    Article Title: Characterization and Recombinant Expression of Terebrid Venom Peptide from Terebra guttata
    Article Snippet: The T4 DNA polymerase-treated insert was annealed into the Xa/LIC vector at 22 °C for 5 min. Then, 7.25 mM EDTA was added, and the components were stirred with a pipet tip for another 5 min at 22 °C. .. The pET-32 Xa/LIC:Tgu6.1 plasmid construct was transformed into E. coli NovaBlue obtained from Novagen. .. Positive clones were screened for via ampicillin and kanamycin resistance.

    Article Title: Proteomic Analysis of and Immune Responses to Ehrlichia chaffeensis Lipoproteins
    Article Snippet: The recombinant plasmids were amplified in E. coli NovaBlue (Novagen). .. Recombinant lipoproteins were expressed in E. coli BL21(DE3).

    Article Title: Equine Arteritis Virus Uses Equine CXCL16 as an Entry Receptor
    Article Snippet: The purified EqCXCL16 fragment was digested with the XhoI and BamHI restriction enzymes and ligated (Rapid DNA ligation kit; Thermo Scientific, Rockford, IL) into the bacterial expression vector pET15b (EMD Millipore Novagen, Temecula, CA) following digestion with the same restriction enzymes. .. The resultant plasmid construct (p15-16A) was used to transform E. coli NovaBlue (EMD Millipore Novagen, Temecula, CA) using a TransformAid kit (Thermo Scientific, Rockford, IL). .. Recombinant plasmid p15-16A (aa 17 to 247) was isolated from ampicillin-resistant clones using a ZR plasmid miniprep kit (Zymo Research, Irvine, CA).

    Real-time Polymerase Chain Reaction:

    Article Title: Phospholipase D from Loxosceleslaeta Spider Venom Induces IL-6, IL-8, CXCL1/GRO-α, and CCL2/MCP-1 Production in Human Skin Fibroblasts and Stimulates Monocytes Migration
    Article Snippet: In addition, each gene was cloned into a cloning vector pCR2.1® -TOPO® using the TOPO TA Cloning® kit (Invitrogen) and transformed into E. coli Novablue (EMD Chemicals-Novagen Brand., Madison, WI, USA) Then, cloned gene plasmids were purified and used to prepare a standard curve for quantitative RT-PCR (RT-qPCR) in real time. .. The real-time efficiencies were determined for each gene from standard curves generated from serial dilutions of plasmids sample.

    Article Title: Integrating artificial with natural cells to translate chemical messages that direct E. coli behaviour
    Article Snippet: Plasmids were amplified in E. coli Novablue (Novagen) and purified with Wizard Plus SV Minipreps DNA Purification System (Promega). .. Transcription–translation reactions used the PURExpress In Vitro Protein Synthesis Kit (New England Biolabs) supplemented with 20 units of Human Placenta RNase Inhibitor (New England Biolabs).

    Incubation:

    Article Title: Characterization and Recombinant Expression of Terebrid Venom Peptide from Terebra guttata
    Article Snippet: The pET-32 Xa/LIC:Tgu6.1 plasmid construct was transformed into E. coli NovaBlue obtained from Novagen. .. Insertion was verified by colony PCR (EMD Millipore) and DNA sequencing.

    Article Title: Phospholipase D from Loxosceleslaeta Spider Venom Induces IL-6, IL-8, CXCL1/GRO-α, and CCL2/MCP-1 Production in Human Skin Fibroblasts and Stimulates Monocytes Migration
    Article Snippet: Thermal cycling conditions were: 95 °C for 3 min, followed by 32 cycles of 95 °C for 30 s, 58 °C for 30 s, and 72 °C for 1 min, and followed by a final incubation at 72 °C for 5 min. Amplification products were separated in 1.5% agarose gel in TAE buffer and stained with EthBr, then gel images were captured in a DNR Bio-Imaging System MF-ChemiBIS 2.0. (Mahale HaHamisha, Jerusalem, Israel) using Gel-Capture software. .. In addition, each gene was cloned into a cloning vector pCR2.1® -TOPO® using the TOPO TA Cloning® kit (Invitrogen) and transformed into E. coli Novablue (EMD Chemicals-Novagen Brand., Madison, WI, USA) Then, cloned gene plasmids were purified and used to prepare a standard curve for quantitative RT-PCR (RT-qPCR) in real time.

    Activity Assay:

    Article Title: A mutant β-glucosidase increases the rate of the cellulose enzymatic hydrolysis
    Article Snippet: The random Sfβgly mutants library was used to transform E. coli NovaBlue(DE3) (Novagen, Darmstadt, Germany). .. The random Sfβgly mutants library was used to transform E. coli NovaBlue(DE3) (Novagen, Darmstadt, Germany).

    Expressing:

    Article Title: A mutant β-glucosidase increases the rate of the cellulose enzymatic hydrolysis
    Article Snippet: 2.1 A sample of the expression vector pCAL coding for the wild-type Sfβgly was submitted to random mutagenesis using the GeneMorph II kit (Agilent Technologies; Santa Clara, US) following the manufacturer's instructions. .. The random Sfβgly mutants library was used to transform E. coli NovaBlue(DE3) (Novagen, Darmstadt, Germany).

    Article Title: Proteomic Analysis of and Immune Responses to Ehrlichia chaffeensis Lipoproteins
    Article Snippet: Paragraph title: Recombinant lipoprotein expression. ... The recombinant plasmids were amplified in E. coli NovaBlue (Novagen).

    Article Title: Characterization of a Novel Serine Protease Inhibitor Gene from a Marine Metagenome
    Article Snippet: The protein expression vector used was pETBlue-2. .. E. coli NovaBlue (Novagen) strain was used as the screening host for the pETBlue-2 bearing inserts.

    Article Title: Display of both N- and C-terminal target fusion proteins on the Aspergillus oryzae cell surface using a chitin-binding module
    Article Snippet: Escherichia coli NovaBlue (Novagen, Inc., Madison, WI) was used as the cloning host for recombinant DNA manipulations. .. Escherichia coli NovaBlue (Novagen, Inc., Madison, WI) was used as the cloning host for recombinant DNA manipulations.

    Article Title: Equine Arteritis Virus Uses Equine CXCL16 as an Entry Receptor
    Article Snippet: Paragraph title: Cloning and expression of EqCXCL16 in Escherichia coli . ... The resultant plasmid construct (p15-16A) was used to transform E. coli NovaBlue (EMD Millipore Novagen, Temecula, CA) using a TransformAid kit (Thermo Scientific, Rockford, IL).

    Article Title: Diversity of hydrolases from hydrothermal vent sediments of the Levante Bay, Vulcano Island (Aeolian archipelago) identified by activity-based metagenomics and biochemical characterization of new esterases and an arabinopyranosidase
    Article Snippet: Paragraph title: Cloning and expression of selected genes in E. coli ... The purified PCR products were then purified, treated with an endonuclease, annealed to His-tag harbouring the pET-46c Ek/LIC vector, and transformed into E. coli NovaBlue according to the protocol of the manufacturer (Novagen, Darmstadt, Germany).

    Article Title: MprAB Regulates the espA Operon in Mycobacterium tuberculosis and Modulates ESX-1 Function and Host Cytokine Response
    Article Snippet: Strains were grown in Middlebrook 7H9 broth containing 0.05% Tween 80 and 10% oleic acid-albumin-dextrose-catalase (OADC) at 37°C with shaking under atmospheric CO2 . .. Escherichia coli NovaBlue and Rosetta(DE3)pLysS (Novagen) were used for general cloning and protein expression, respectively. .. Briefly, M. tuberculosis strains were grown to mid-exponential phase and total RNA was extracted by bead beating, followed by further purification and DNase treatment.

    Article Title: The ?-propeller gene Rv1057 of Mycobacterium tuberculosis has a complex promoter directly regulated by both the MprAB and TrcRS two-component systems
    Article Snippet: M. tuberculosis strain Rv-D981 is an mprAB deletion mutant derived from the laboratory strain H37Rv, and Rv-D981Com is the mprAB -complemented strain generated from Rv-D981 . .. Escherichia coli Novablue and Rosetta(DE3)pLysS (Novagen) were used for general cloning purposes and protein expression, respectively. .. Rv1057 promoter fusions were constructed in the integrating vector pSM128 by inserting sections of the Rv1056-Rv1057 intergenic region upstream of lacZ .

    Transformation Assay:

    Article Title: Penicillin-Binding Protein of Ehrlichia chaffeensis: Cytokine Induction Through MyD88-Dependent Pathway
    Article Snippet: The amplified fragments were digested with restriction enzymes and ligated into restriction enzyme–digested pET-33b(+) (Novagen). .. Escherichia coli Novablue (Novagen) was transformed and the plasmids were extracted. .. The cloned fragments were confirmed by DNA sequencing.

    Article Title: Characterization and Recombinant Expression of Terebrid Venom Peptide from Terebra guttata
    Article Snippet: The T4 DNA polymerase-treated insert was annealed into the Xa/LIC vector at 22 °C for 5 min. Then, 7.25 mM EDTA was added, and the components were stirred with a pipet tip for another 5 min at 22 °C. .. The pET-32 Xa/LIC:Tgu6.1 plasmid construct was transformed into E. coli NovaBlue obtained from Novagen. .. Positive clones were screened for via ampicillin and kanamycin resistance.

    Article Title: Phospholipase D from Loxosceleslaeta Spider Venom Induces IL-6, IL-8, CXCL1/GRO-α, and CCL2/MCP-1 Production in Human Skin Fibroblasts and Stimulates Monocytes Migration
    Article Snippet: Thermal cycling conditions were: 95 °C for 3 min, followed by 32 cycles of 95 °C for 30 s, 58 °C for 30 s, and 72 °C for 1 min, and followed by a final incubation at 72 °C for 5 min. Amplification products were separated in 1.5% agarose gel in TAE buffer and stained with EthBr, then gel images were captured in a DNR Bio-Imaging System MF-ChemiBIS 2.0. (Mahale HaHamisha, Jerusalem, Israel) using Gel-Capture software. .. In addition, each gene was cloned into a cloning vector pCR2.1® -TOPO® using the TOPO TA Cloning® kit (Invitrogen) and transformed into E. coli Novablue (EMD Chemicals-Novagen Brand., Madison, WI, USA) Then, cloned gene plasmids were purified and used to prepare a standard curve for quantitative RT-PCR (RT-qPCR) in real time. .. The real-time efficiencies were determined for each gene from standard curves generated from serial dilutions of plasmids sample.

    Article Title: Proteomic Analysis of and Immune Responses to Ehrlichia chaffeensis Lipoproteins
    Article Snippet: The recombinant plasmids were amplified in E. coli NovaBlue (Novagen). .. Recombinant lipoproteins were expressed in E. coli BL21(DE3).

    Article Title: Equine Arteritis Virus Uses Equine CXCL16 as an Entry Receptor
    Article Snippet: The resultant plasmid construct (p15-16A) was used to transform E. coli NovaBlue (EMD Millipore Novagen, Temecula, CA) using a TransformAid kit (Thermo Scientific, Rockford, IL). .. Recombinant plasmid p15-16A (aa 17 to 247) was isolated from ampicillin-resistant clones using a ZR plasmid miniprep kit (Zymo Research, Irvine, CA).

    Article Title: Diversity of hydrolases from hydrothermal vent sediments of the Levante Bay, Vulcano Island (Aeolian archipelago) identified by activity-based metagenomics and biochemical characterization of new esterases and an arabinopyranosidase
    Article Snippet: The reactions were done in a Techne TC-5000 cycler (CA, USA) using the following programme: 1 cycle of 95 °C/3 min following 35 cycles of 95 °C for 30s, 55 °C for 30s, and 72 °C for 1 min per 1000 nucleotides, followed by one extension at 72 °C for 5 min. .. The purified PCR products were then purified, treated with an endonuclease, annealed to His-tag harbouring the pET-46c Ek/LIC vector, and transformed into E. coli NovaBlue according to the protocol of the manufacturer (Novagen, Darmstadt, Germany). .. The DNA inserts in the resulting plasmids were verified by sequencing services of Macrogen Ltd. (Amsterdam, The Netherlands).

    Derivative Assay:

    Article Title: Display of both N- and C-terminal target fusion proteins on the Aspergillus oryzae cell surface using a chitin-binding module
    Article Snippet: Escherichia coli NovaBlue (Novagen, Inc., Madison, WI) was used as the cloning host for recombinant DNA manipulations. .. Escherichia coli NovaBlue (Novagen, Inc., Madison, WI) was used as the cloning host for recombinant DNA manipulations.

    Article Title: The ?-propeller gene Rv1057 of Mycobacterium tuberculosis has a complex promoter directly regulated by both the MprAB and TrcRS two-component systems
    Article Snippet: M. tuberculosis strain Rv-D981 is an mprAB deletion mutant derived from the laboratory strain H37Rv, and Rv-D981Com is the mprAB -complemented strain generated from Rv-D981 . .. Escherichia coli Novablue and Rosetta(DE3)pLysS (Novagen) were used for general cloning purposes and protein expression, respectively.

    Conjugation Assay:

    Article Title: Disruption of poly (3-hydroxyalkanoate) depolymerase gene and overexpression of three poly (3-hydroxybutyrate) biosynthetic genes improve poly (3-hydroxybutyrate) production from nitrogen rich medium by Rhodobacter sphaeroides
    Article Snippet: For PHB production, R. sphaeroides HJ derivative strains were grown in AAY medium consisting basal medium, 100 mM sodium acetate, 1.0 g l−1 yeast extract, and 10 mM AS. .. Escherichia coli NovaBlue (Novagen, Darmstadt, Germany) and S17-1 strains were used for DNA manipulation and conjugation transfer, respectively. .. E. coli strains were aerobically grown at 37 °C in Luria-Bertani (LB) medium (10 g l−1 tryptone, 5 g l−1 yeast extract, and 10 g l−1 sodium chloride).

    Ligation:

    Article Title: Characterization and Recombinant Expression of Terebrid Venom Peptide from Terebra guttata
    Article Snippet: The gene insert was amplified using primers 5′-GGTATTGAGGGTCGCATATTATATTATTTA-3′ and 5′-AGAGGAGAGTTAGAGCCATAATAATATTTA-3′ (IDTDNA), which contained the requisite ligation-independent cloning (LIC); overhangs underlined in the sequence above. .. The pET-32 Xa/LIC:Tgu6.1 plasmid construct was transformed into E. coli NovaBlue obtained from Novagen.

    Article Title: Diversity of hydrolases from hydrothermal vent sediments of the Levante Bay, Vulcano Island (Aeolian archipelago) identified by activity-based metagenomics and biochemical characterization of new esterases and an arabinopyranosidase
    Article Snippet: The PCR primer pairs (Table ) with the nucleotide adapters used in this research were designed following a ligation-independent cloning protocol of Novagen (Darmstadt, Germany). .. The purified PCR products were then purified, treated with an endonuclease, annealed to His-tag harbouring the pET-46c Ek/LIC vector, and transformed into E. coli NovaBlue according to the protocol of the manufacturer (Novagen, Darmstadt, Germany).

    Library Screening:

    Article Title: A mutant β-glucosidase increases the rate of the cellulose enzymatic hydrolysis
    Article Snippet: Paragraph title: Random mutagenesis and library screening ... The random Sfβgly mutants library was used to transform E. coli NovaBlue(DE3) (Novagen, Darmstadt, Germany).

    Generated:

    Article Title: The ?-propeller gene Rv1057 of Mycobacterium tuberculosis has a complex promoter directly regulated by both the MprAB and TrcRS two-component systems
    Article Snippet: M. tuberculosis strain Rv-D981 is an mprAB deletion mutant derived from the laboratory strain H37Rv, and Rv-D981Com is the mprAB -complemented strain generated from Rv-D981 . .. Escherichia coli Novablue and Rosetta(DE3)pLysS (Novagen) were used for general cloning purposes and protein expression, respectively.

    other:

    Article Title: Mycobacterium tuberculosis Rv1096 protein: gene cloning, protein expression, and peptidoglycan deacetylase activity
    Article Snippet: E. coli NovaBlue (Novagen, Madison, WI) and ER2566 (Novagen) strains were routinely grown in Luria-Bertani media (LB, Invitrogen, Carlsbad, CA).

    DNA Sequencing:

    Article Title: Characterization and Recombinant Expression of Terebrid Venom Peptide from Terebra guttata
    Article Snippet: The pET-32 Xa/LIC:Tgu6.1 plasmid construct was transformed into E. coli NovaBlue obtained from Novagen. .. The pET-32 Xa/LIC:Tgu6.1 plasmid construct was transformed into E. coli NovaBlue obtained from Novagen.

    Sequencing:

    Article Title: Characterization and Recombinant Expression of Terebrid Venom Peptide from Terebra guttata
    Article Snippet: The gene insert was amplified using primers 5′-GGTATTGAGGGTCGCATATTATATTATTTA-3′ and 5′-AGAGGAGAGTTAGAGCCATAATAATATTTA-3′ (IDTDNA), which contained the requisite ligation-independent cloning (LIC); overhangs underlined in the sequence above. .. The pET-32 Xa/LIC:Tgu6.1 plasmid construct was transformed into E. coli NovaBlue obtained from Novagen.

    Article Title: Phospholipase D from Loxosceleslaeta Spider Venom Induces IL-6, IL-8, CXCL1/GRO-α, and CCL2/MCP-1 Production in Human Skin Fibroblasts and Stimulates Monocytes Migration
    Article Snippet: The sequence of primers was as follows: IL-6 (sense) 5′-CTGCTCCTGGTGTTGCCT-3′, IL-6 (antisense) 5′-CTGAAGAGGTGAGTGGCTGT-3′; IL-8 (sense) 5′-ACTGAGAGTGATTGAGAGTGG-3′, IL-8 (antisense) 5′-AGTTTTCCTTGGGGTCCAGA-3′; CXCL1 (sense) 5′-GACCAGAAGGGAGGAGGAAG-3′, CXCL1 (antisense) 5′-ATGCTCAAACACATTAGGCACA-3′; CCL2 (sense) 5′-AAGCAGAAGTGGGTTCAGGATT-3′, CCL2 (antisense) 5′-TTGGGTTGTGGAGTGAGTGTT-3′; Beta actin (sense) 5′-GTTGCGTTACACCCTTTCTTG-3′, Beta actin (antisense) 5′-CACCTTCACCGTTCCAGTTT-3’. .. In addition, each gene was cloned into a cloning vector pCR2.1® -TOPO® using the TOPO TA Cloning® kit (Invitrogen) and transformed into E. coli Novablue (EMD Chemicals-Novagen Brand., Madison, WI, USA) Then, cloned gene plasmids were purified and used to prepare a standard curve for quantitative RT-PCR (RT-qPCR) in real time.

    Article Title: Proteomic Analysis of and Immune Responses to Ehrlichia chaffeensis Lipoproteins
    Article Snippet: Table S1 in the supplemental material shows primers flanked with an NdeI site on the forward primer and an XhoI site on the reverse primer, covering the predicted mature sequence (without the signal peptide) for lipoprotein ECH_0482 and full-length ORFs (including the signal peptide) for ECH_0558 and ECH_0929. .. The recombinant plasmids were amplified in E. coli NovaBlue (Novagen).

    Article Title: Equine Arteritis Virus Uses Equine CXCL16 as an Entry Receptor
    Article Snippet: The sequence encoding the entire EqCXCL16 without the first 16 amino acid residues from the signal sequences was amplified from cDNA made from mRNA extracted from equine monocytes using a Smart cDNA synthesis kit (Clontech Laboratories Inc., Mountain View, CA). .. The resultant plasmid construct (p15-16A) was used to transform E. coli NovaBlue (EMD Millipore Novagen, Temecula, CA) using a TransformAid kit (Thermo Scientific, Rockford, IL).

    Sonication:

    Article Title: Equine Arteritis Virus Uses Equine CXCL16 as an Entry Receptor
    Article Snippet: The resultant plasmid construct (p15-16A) was used to transform E. coli NovaBlue (EMD Millipore Novagen, Temecula, CA) using a TransformAid kit (Thermo Scientific, Rockford, IL). .. The resultant plasmid construct (p15-16A) was used to transform E. coli NovaBlue (EMD Millipore Novagen, Temecula, CA) using a TransformAid kit (Thermo Scientific, Rockford, IL).

    Affinity Purification:

    Article Title: Proteomic Analysis of and Immune Responses to Ehrlichia chaffeensis Lipoproteins
    Article Snippet: The recombinant plasmids were amplified in E. coli NovaBlue (Novagen). .. When the optical density at 600 nm of the E. coli culture reached 0.6, isopropyl-β- d -thiogalactopyranoside (Gold Bio Technology, St. Louis, MO) was added at a final concentration of 1 mM.

    Recombinant:

    Article Title: Disruption of PHO13 improves ethanol production via the xylose isomerase pathway
    Article Snippet: The effect of PHO13 disruption on the metabolome was investigated by both transcriptome and metabolite analysis. .. Escherichia coli NovaBlue (Novagen, Inc., Madison, WI, USA) was used as the host strain for recombinant DNA manipulation and was grown in Luria–Bertani medium (10 g/L tryptone, 5 g/L yeast extract, and 5 g/L sodium chloride) containing 100 mg/L ampicillin. .. S. cerevisiae YPH499 (MAT a ade2 his3 leu2 lys2 trp1 ura3 , purchased from Stratagene, La Jolla, CA, USA) was used as the host strain.

    Article Title: Development of a GIN11/FRT-based multiple-gene integration technique affording inhibitor-tolerant, hemicellulolytic, xylose-utilizing abilities to industrial Saccharomyces cerevisiae strains for ethanol production from undetoxified lignocellulosic hemicelluloses
    Article Snippet: Yeast strains were routinely cultivated at 30°C in synthetic medium [SD medium; 6.7 g/L of yeast nitrogen base without amino acids (Difco Laboratories, Detroit, MI), 20 g/L of glucose] supplemented with antibiotics, and in YPD medium (20 g/L peptone, 10 g/L yeast extract, 20 g/L glucose). .. Escherichia coli NovaBlue (Novagen, Inc., Madison, WI) was used as the host strain for recombinant DNA manipulation. .. E. coli was grown in Luria-Bertani medium (10 g/L peptone, 5 g/L yeast extract, and 5 g/L sodium chloride) containing 100 mg/L ampicillin.

    Article Title: Aspergillus oryzae-based cell factory for direct kojic acid production from cellulose
    Article Snippet: The washed mycelium was recultivated in 500 mL Erlenmeyer flasks with baffles containing 100 mL CY medium at 30°C that were shaken at 150 rpm using an orbital shaker. .. E. coli NovaBlue (Novagen, Inc., Madison, WI, USA) was used as the cloning host for recombinant DNA manipulations. .. The bacterium was grown in Luria–Bertani medium (1% tryptone, 0.5% yeast extract, and 0.5% NaCl) containing 0.1 mg/mL of ampicillin.

    Article Title: Penicillin-Binding Protein of Ehrlichia chaffeensis: Cytokine Induction Through MyD88-Dependent Pathway
    Article Snippet: Paragraph title: Recombinant Proteins ... Escherichia coli Novablue (Novagen) was transformed and the plasmids were extracted.

    Article Title: Characterization and Recombinant Expression of Terebrid Venom Peptide from Terebra guttata
    Article Snippet: Paragraph title: 3.1. Construction of Recombinant Plasmid ... The pET-32 Xa/LIC:Tgu6.1 plasmid construct was transformed into E. coli NovaBlue obtained from Novagen.

    Article Title: Proteomic Analysis of and Immune Responses to Ehrlichia chaffeensis Lipoproteins
    Article Snippet: Inserts excised from pCR-Blunt II were directionally subcloned into E. coli expression vector pET29a(+) (Novagen, San Diego, CA) digested with NdeI and XhoI. .. The recombinant plasmids were amplified in E. coli NovaBlue (Novagen). .. Recombinant lipoproteins were expressed in E. coli BL21(DE3).

    Article Title: Metabolic pathway engineering based on metabolomics confers acetic and formic acid tolerance to a recombinant xylose-fermenting strain of Saccharomyces cerevisiae
    Article Snippet: Yeast strains were routinely cultivated at 30°C in synthetic medium [SD medium; 6.7 g/L of yeast nitrogen base without amino acids (Difco Laboratories, Detroit, MI), 20 g/L of glucose] supplemented with appropriate amino acids and nucleotides, and in YPD medium (20 g/L peptone, 10 g/L yeast extract, 20 g/L glucose). .. Escherichia coli NovaBlue (Novagen, Inc., Madison, WI) was used as the host strain for recombinant DNA manipulation. .. E. coli was grown in Luria-Bertani medium (10 g/L peptone, 5 g/L yeast extract, and 5 g/L sodium chloride) containing 100 mg/L ampicillin.

    Article Title: Display of both N- and C-terminal target fusion proteins on the Aspergillus oryzae cell surface using a chitin-binding module
    Article Snippet: Using CBM as an anchor protein, we tested the displaying possibilities of green fluorescent protein (GFP) and triacylglycerol lipase from A. oryzae (tglA). .. Escherichia coli NovaBlue (Novagen, Inc., Madison, WI) was used as the cloning host for recombinant DNA manipulations. .. The bacterium was grown in Luria–Bertani medium (1% tryptone, 0.5% yeast extract, and 0.5% NaCl) containing 0.1 mg/ml of ampicillin.

    Article Title: Equine Arteritis Virus Uses Equine CXCL16 as an Entry Receptor
    Article Snippet: The resultant plasmid construct (p15-16A) was used to transform E. coli NovaBlue (EMD Millipore Novagen, Temecula, CA) using a TransformAid kit (Thermo Scientific, Rockford, IL). .. Recombinant plasmid p15-16A (aa 17 to 247) was isolated from ampicillin-resistant clones using a ZR plasmid miniprep kit (Zymo Research, Irvine, CA).

    Fluorescence:

    Article Title: Integrating artificial with natural cells to translate chemical messages that direct E. coli behaviour
    Article Snippet: Plasmids were amplified in E. coli Novablue (Novagen) and purified with Wizard Plus SV Minipreps DNA Purification System (Promega). .. Transcription–translation reactions used the PURExpress In Vitro Protein Synthesis Kit (New England Biolabs) supplemented with 20 units of Human Placenta RNase Inhibitor (New England Biolabs).

    Electron Microscopy:

    Article Title: Treponema denticola PrcB Is Required for Expression and Activity of the PrcA-PrtP (Dentilisin) Complex
    Article Snippet: T. denticola ATCC 35405, ATCC 33520, and OTK and isogenic mutants of 35405 (Table ) were grown in NOS broth or NOS/GN semisolid medium as previously described ( , ), with erythromycin (Em) (40 μg ml−1 ) added as appropriate. .. Escherichia coli NovaBlue (Novagen, Inc., Madison, WI) and JM109 ( ) were used as hosts for cloning.

    Mutagenesis:

    Article Title: A mutant β-glucosidase increases the rate of the cellulose enzymatic hydrolysis
    Article Snippet: Paragraph title: Random mutagenesis and library screening ... The random Sfβgly mutants library was used to transform E. coli NovaBlue(DE3) (Novagen, Darmstadt, Germany).

    Article Title: Display of both N- and C-terminal target fusion proteins on the Aspergillus oryzae cell surface using a chitin-binding module
    Article Snippet: Escherichia coli NovaBlue (Novagen, Inc., Madison, WI) was used as the cloning host for recombinant DNA manipulations. .. Escherichia coli NovaBlue (Novagen, Inc., Madison, WI) was used as the cloning host for recombinant DNA manipulations.

    Article Title: MprAB Regulates the espA Operon in Mycobacterium tuberculosis and Modulates ESX-1 Function and Host Cytokine Response
    Article Snippet: ST100 is a phoP mutant of H37Rv ( ). .. Escherichia coli NovaBlue and Rosetta(DE3)pLysS (Novagen) were used for general cloning and protein expression, respectively.

    Article Title: The ?-propeller gene Rv1057 of Mycobacterium tuberculosis has a complex promoter directly regulated by both the MprAB and TrcRS two-component systems
    Article Snippet: M. tuberculosis strain Rv-D981 is an mprAB deletion mutant derived from the laboratory strain H37Rv, and Rv-D981Com is the mprAB -complemented strain generated from Rv-D981 . .. Escherichia coli Novablue and Rosetta(DE3)pLysS (Novagen) were used for general cloning purposes and protein expression, respectively.

    Isolation:

    Article Title: Disruption of poly (3-hydroxyalkanoate) depolymerase gene and overexpression of three poly (3-hydroxybutyrate) biosynthetic genes improve poly (3-hydroxybutyrate) production from nitrogen rich medium by Rhodobacter sphaeroides
    Article Snippet: R. sphaeroides HJ, which was isolated in a previous study [ ], and R. sphaeroides 2.4.1 were used as a host strain and a source of genes, respectively. .. Escherichia coli NovaBlue (Novagen, Darmstadt, Germany) and S17-1 strains were used for DNA manipulation and conjugation transfer, respectively.

    Microscopy:

    Article Title: Treponema denticola PrcB Is Required for Expression and Activity of the PrcA-PrtP (Dentilisin) Complex
    Article Snippet: Cultures were examined by phase-contrast microscopy for purity and typical strain morphology before use. .. Escherichia coli NovaBlue (Novagen, Inc., Madison, WI) and JM109 ( ) were used as hosts for cloning.

    Purification:

    Article Title: Penicillin-Binding Protein of Ehrlichia chaffeensis: Cytokine Induction Through MyD88-Dependent Pathway
    Article Snippet: Escherichia coli Novablue (Novagen) was transformed and the plasmids were extracted. .. Escherichia coli BL21 (DE3) (Novagen) cells were transformed with the plasmid and induced to express the recombinant proteins with isopropyl-thio-ß- d -galactoside (Gold Biotechnology).

    Article Title: Characterization and Recombinant Expression of Terebrid Venom Peptide from Terebra guttata
    Article Snippet: The PCR amplified insert was purified with SpinPrep Gel DNA Kit (Novagen, Darmstadt, Germany) and cloned via LIC into vector pET-32 Xa/LIC (EMD Millipore, Darmstadt, Germany). .. The pET-32 Xa/LIC:Tgu6.1 plasmid construct was transformed into E. coli NovaBlue obtained from Novagen.

    Article Title: Phospholipase D from Loxosceleslaeta Spider Venom Induces IL-6, IL-8, CXCL1/GRO-α, and CCL2/MCP-1 Production in Human Skin Fibroblasts and Stimulates Monocytes Migration
    Article Snippet: Thermal cycling conditions were: 95 °C for 3 min, followed by 32 cycles of 95 °C for 30 s, 58 °C for 30 s, and 72 °C for 1 min, and followed by a final incubation at 72 °C for 5 min. Amplification products were separated in 1.5% agarose gel in TAE buffer and stained with EthBr, then gel images were captured in a DNR Bio-Imaging System MF-ChemiBIS 2.0. (Mahale HaHamisha, Jerusalem, Israel) using Gel-Capture software. .. In addition, each gene was cloned into a cloning vector pCR2.1® -TOPO® using the TOPO TA Cloning® kit (Invitrogen) and transformed into E. coli Novablue (EMD Chemicals-Novagen Brand., Madison, WI, USA) Then, cloned gene plasmids were purified and used to prepare a standard curve for quantitative RT-PCR (RT-qPCR) in real time. .. The real-time efficiencies were determined for each gene from standard curves generated from serial dilutions of plasmids sample.

    Article Title: Integrating artificial with natural cells to translate chemical messages that direct E. coli behaviour
    Article Snippet: Data are reported as averages with standard error, or representative runs are shown. .. Plasmids were amplified in E. coli Novablue (Novagen) and purified with Wizard Plus SV Minipreps DNA Purification System (Promega). .. Plasmid DNA was phenol–chloroform extracted, ethanol precipitated and resuspended in deionized and diethyl pyrocarbonate-treated water.

    Article Title: Proteomic Analysis of and Immune Responses to Ehrlichia chaffeensis Lipoproteins
    Article Snippet: With these primers, lipoprotein DNA fragments were amplified from purified E. chaffeensis DNA using Pfu Turbo polymerase. .. The recombinant plasmids were amplified in E. coli NovaBlue (Novagen).

    Article Title: Equine Arteritis Virus Uses Equine CXCL16 as an Entry Receptor
    Article Snippet: The purified EqCXCL16 fragment was digested with the XhoI and BamHI restriction enzymes and ligated (Rapid DNA ligation kit; Thermo Scientific, Rockford, IL) into the bacterial expression vector pET15b (EMD Millipore Novagen, Temecula, CA) following digestion with the same restriction enzymes. .. The resultant plasmid construct (p15-16A) was used to transform E. coli NovaBlue (EMD Millipore Novagen, Temecula, CA) using a TransformAid kit (Thermo Scientific, Rockford, IL).

    Article Title: Diversity of hydrolases from hydrothermal vent sediments of the Levante Bay, Vulcano Island (Aeolian archipelago) identified by activity-based metagenomics and biochemical characterization of new esterases and an arabinopyranosidase
    Article Snippet: The reactions were done in a Techne TC-5000 cycler (CA, USA) using the following programme: 1 cycle of 95 °C/3 min following 35 cycles of 95 °C for 30s, 55 °C for 30s, and 72 °C for 1 min per 1000 nucleotides, followed by one extension at 72 °C for 5 min. .. The purified PCR products were then purified, treated with an endonuclease, annealed to His-tag harbouring the pET-46c Ek/LIC vector, and transformed into E. coli NovaBlue according to the protocol of the manufacturer (Novagen, Darmstadt, Germany). .. The DNA inserts in the resulting plasmids were verified by sequencing services of Macrogen Ltd. (Amsterdam, The Netherlands).

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: Phospholipase D from Loxosceleslaeta Spider Venom Induces IL-6, IL-8, CXCL1/GRO-α, and CCL2/MCP-1 Production in Human Skin Fibroblasts and Stimulates Monocytes Migration
    Article Snippet: Paragraph title: 4.6. RT-PCR and Quantitative Real-Time RT-PCR (RT-qPCR) ... In addition, each gene was cloned into a cloning vector pCR2.1® -TOPO® using the TOPO TA Cloning® kit (Invitrogen) and transformed into E. coli Novablue (EMD Chemicals-Novagen Brand., Madison, WI, USA) Then, cloned gene plasmids were purified and used to prepare a standard curve for quantitative RT-PCR (RT-qPCR) in real time.

    Quantitative RT-PCR:

    Article Title: Phospholipase D from Loxosceleslaeta Spider Venom Induces IL-6, IL-8, CXCL1/GRO-α, and CCL2/MCP-1 Production in Human Skin Fibroblasts and Stimulates Monocytes Migration
    Article Snippet: Thermal cycling conditions were: 95 °C for 3 min, followed by 32 cycles of 95 °C for 30 s, 58 °C for 30 s, and 72 °C for 1 min, and followed by a final incubation at 72 °C for 5 min. Amplification products were separated in 1.5% agarose gel in TAE buffer and stained with EthBr, then gel images were captured in a DNR Bio-Imaging System MF-ChemiBIS 2.0. (Mahale HaHamisha, Jerusalem, Israel) using Gel-Capture software. .. In addition, each gene was cloned into a cloning vector pCR2.1® -TOPO® using the TOPO TA Cloning® kit (Invitrogen) and transformed into E. coli Novablue (EMD Chemicals-Novagen Brand., Madison, WI, USA) Then, cloned gene plasmids were purified and used to prepare a standard curve for quantitative RT-PCR (RT-qPCR) in real time. .. The real-time efficiencies were determined for each gene from standard curves generated from serial dilutions of plasmids sample.

    IA:

    Article Title: Equine Arteritis Virus Uses Equine CXCL16 as an Entry Receptor
    Article Snippet: The cDNA amplification was carried out according to a standard laboratory PCR protocol using forward primer cx16-15F (5′-GCG CTCGAG GCGTTGCTGACTCTGCAAGG-3′) and reverse primer cx16-15R (5′-GC GGATCC GCACTGCCACTGTAACTGAT-3′) (IDT, Coralville, IA). .. The resultant plasmid construct (p15-16A) was used to transform E. coli NovaBlue (EMD Millipore Novagen, Temecula, CA) using a TransformAid kit (Thermo Scientific, Rockford, IL).

    Plasmid Preparation:

    Article Title: A mutant β-glucosidase increases the rate of the cellulose enzymatic hydrolysis
    Article Snippet: 2.1 A sample of the expression vector pCAL coding for the wild-type Sfβgly was submitted to random mutagenesis using the GeneMorph II kit (Agilent Technologies; Santa Clara, US) following the manufacturer's instructions. .. The random Sfβgly mutants library was used to transform E. coli NovaBlue(DE3) (Novagen, Darmstadt, Germany).

    Article Title: Penicillin-Binding Protein of Ehrlichia chaffeensis: Cytokine Induction Through MyD88-Dependent Pathway
    Article Snippet: Escherichia coli Novablue (Novagen) was transformed and the plasmids were extracted. .. Escherichia coli Novablue (Novagen) was transformed and the plasmids were extracted.

    Article Title: Treponema denticola PrcB Is Required for Expression and Activity of the PrcA-PrtP (Dentilisin) Complex
    Article Snippet: Escherichia coli NovaBlue (Novagen, Inc., Madison, WI) and JM109 ( ) were used as hosts for cloning. .. E. coli was grown in LB agar or broth medium with ampicillin (50 μg ml−1 ), kanamycin (30 μg ml−1 ), and Em (200 μg ml−1 ), as appropriate.

    Article Title: Characterization and Recombinant Expression of Terebrid Venom Peptide from Terebra guttata
    Article Snippet: The T4 DNA polymerase-treated insert was annealed into the Xa/LIC vector at 22 °C for 5 min. Then, 7.25 mM EDTA was added, and the components were stirred with a pipet tip for another 5 min at 22 °C. .. The pET-32 Xa/LIC:Tgu6.1 plasmid construct was transformed into E. coli NovaBlue obtained from Novagen. .. Positive clones were screened for via ampicillin and kanamycin resistance.

    Article Title: Phospholipase D from Loxosceleslaeta Spider Venom Induces IL-6, IL-8, CXCL1/GRO-α, and CCL2/MCP-1 Production in Human Skin Fibroblasts and Stimulates Monocytes Migration
    Article Snippet: Thermal cycling conditions were: 95 °C for 3 min, followed by 32 cycles of 95 °C for 30 s, 58 °C for 30 s, and 72 °C for 1 min, and followed by a final incubation at 72 °C for 5 min. Amplification products were separated in 1.5% agarose gel in TAE buffer and stained with EthBr, then gel images were captured in a DNR Bio-Imaging System MF-ChemiBIS 2.0. (Mahale HaHamisha, Jerusalem, Israel) using Gel-Capture software. .. In addition, each gene was cloned into a cloning vector pCR2.1® -TOPO® using the TOPO TA Cloning® kit (Invitrogen) and transformed into E. coli Novablue (EMD Chemicals-Novagen Brand., Madison, WI, USA) Then, cloned gene plasmids were purified and used to prepare a standard curve for quantitative RT-PCR (RT-qPCR) in real time. .. The real-time efficiencies were determined for each gene from standard curves generated from serial dilutions of plasmids sample.

    Article Title: Proteomic Analysis of and Immune Responses to Ehrlichia chaffeensis Lipoproteins
    Article Snippet: Inserts excised from pCR-Blunt II were directionally subcloned into E. coli expression vector pET29a(+) (Novagen, San Diego, CA) digested with NdeI and XhoI. .. The recombinant plasmids were amplified in E. coli NovaBlue (Novagen).

    Article Title: Characterization of a Novel Serine Protease Inhibitor Gene from a Marine Metagenome
    Article Snippet: Paragraph title: 3.1. Plasmid Vectors, Bacterium Strains, and Growth Conditions ... E. coli NovaBlue (Novagen) strain was used as the screening host for the pETBlue-2 bearing inserts.

    Article Title: Equine Arteritis Virus Uses Equine CXCL16 as an Entry Receptor
    Article Snippet: The purified EqCXCL16 fragment was digested with the XhoI and BamHI restriction enzymes and ligated (Rapid DNA ligation kit; Thermo Scientific, Rockford, IL) into the bacterial expression vector pET15b (EMD Millipore Novagen, Temecula, CA) following digestion with the same restriction enzymes. .. The resultant plasmid construct (p15-16A) was used to transform E. coli NovaBlue (EMD Millipore Novagen, Temecula, CA) using a TransformAid kit (Thermo Scientific, Rockford, IL). .. Recombinant plasmid p15-16A (aa 17 to 247) was isolated from ampicillin-resistant clones using a ZR plasmid miniprep kit (Zymo Research, Irvine, CA).

    Article Title: Diversity of hydrolases from hydrothermal vent sediments of the Levante Bay, Vulcano Island (Aeolian archipelago) identified by activity-based metagenomics and biochemical characterization of new esterases and an arabinopyranosidase
    Article Snippet: The reactions were done in a Techne TC-5000 cycler (CA, USA) using the following programme: 1 cycle of 95 °C/3 min following 35 cycles of 95 °C for 30s, 55 °C for 30s, and 72 °C for 1 min per 1000 nucleotides, followed by one extension at 72 °C for 5 min. .. The purified PCR products were then purified, treated with an endonuclease, annealed to His-tag harbouring the pET-46c Ek/LIC vector, and transformed into E. coli NovaBlue according to the protocol of the manufacturer (Novagen, Darmstadt, Germany). .. The DNA inserts in the resulting plasmids were verified by sequencing services of Macrogen Ltd. (Amsterdam, The Netherlands).

    Software:

    Article Title: Phospholipase D from Loxosceleslaeta Spider Venom Induces IL-6, IL-8, CXCL1/GRO-α, and CCL2/MCP-1 Production in Human Skin Fibroblasts and Stimulates Monocytes Migration
    Article Snippet: Thermal cycling conditions were: 95 °C for 3 min, followed by 32 cycles of 95 °C for 30 s, 58 °C for 30 s, and 72 °C for 1 min, and followed by a final incubation at 72 °C for 5 min. Amplification products were separated in 1.5% agarose gel in TAE buffer and stained with EthBr, then gel images were captured in a DNR Bio-Imaging System MF-ChemiBIS 2.0. (Mahale HaHamisha, Jerusalem, Israel) using Gel-Capture software. .. In addition, each gene was cloned into a cloning vector pCR2.1® -TOPO® using the TOPO TA Cloning® kit (Invitrogen) and transformed into E. coli Novablue (EMD Chemicals-Novagen Brand., Madison, WI, USA) Then, cloned gene plasmids were purified and used to prepare a standard curve for quantitative RT-PCR (RT-qPCR) in real time.

    SYBR Green Assay:

    Article Title: Phospholipase D from Loxosceleslaeta Spider Venom Induces IL-6, IL-8, CXCL1/GRO-α, and CCL2/MCP-1 Production in Human Skin Fibroblasts and Stimulates Monocytes Migration
    Article Snippet: In addition, each gene was cloned into a cloning vector pCR2.1® -TOPO® using the TOPO TA Cloning® kit (Invitrogen) and transformed into E. coli Novablue (EMD Chemicals-Novagen Brand., Madison, WI, USA) Then, cloned gene plasmids were purified and used to prepare a standard curve for quantitative RT-PCR (RT-qPCR) in real time. .. Then, RT-qPCR was then performed using a thermal cycler StepOnePlus™ Real-Time PCR System (Applied Biosystems., Foster City, CA, USA).

    Article Title: Integrating artificial with natural cells to translate chemical messages that direct E. coli behaviour
    Article Snippet: Plasmids were amplified in E. coli Novablue (Novagen) and purified with Wizard Plus SV Minipreps DNA Purification System (Promega). .. Transcription–translation reactions used the PURExpress In Vitro Protein Synthesis Kit (New England Biolabs) supplemented with 20 units of Human Placenta RNase Inhibitor (New England Biolabs).

    Positron Emission Tomography:

    Article Title: Penicillin-Binding Protein of Ehrlichia chaffeensis: Cytokine Induction Through MyD88-Dependent Pathway
    Article Snippet: The amplified fragments were digested with restriction enzymes and ligated into restriction enzyme–digested pET-33b(+) (Novagen). .. Escherichia coli Novablue (Novagen) was transformed and the plasmids were extracted.

    Article Title: Characterization and Recombinant Expression of Terebrid Venom Peptide from Terebra guttata
    Article Snippet: The T4 DNA polymerase-treated insert was annealed into the Xa/LIC vector at 22 °C for 5 min. Then, 7.25 mM EDTA was added, and the components were stirred with a pipet tip for another 5 min at 22 °C. .. The pET-32 Xa/LIC:Tgu6.1 plasmid construct was transformed into E. coli NovaBlue obtained from Novagen. .. Positive clones were screened for via ampicillin and kanamycin resistance.

    Article Title: Diversity of hydrolases from hydrothermal vent sediments of the Levante Bay, Vulcano Island (Aeolian archipelago) identified by activity-based metagenomics and biochemical characterization of new esterases and an arabinopyranosidase
    Article Snippet: The reactions were done in a Techne TC-5000 cycler (CA, USA) using the following programme: 1 cycle of 95 °C/3 min following 35 cycles of 95 °C for 30s, 55 °C for 30s, and 72 °C for 1 min per 1000 nucleotides, followed by one extension at 72 °C for 5 min. .. The purified PCR products were then purified, treated with an endonuclease, annealed to His-tag harbouring the pET-46c Ek/LIC vector, and transformed into E. coli NovaBlue according to the protocol of the manufacturer (Novagen, Darmstadt, Germany). .. The DNA inserts in the resulting plasmids were verified by sequencing services of Macrogen Ltd. (Amsterdam, The Netherlands).

    Agarose Gel Electrophoresis:

    Article Title: Phospholipase D from Loxosceleslaeta Spider Venom Induces IL-6, IL-8, CXCL1/GRO-α, and CCL2/MCP-1 Production in Human Skin Fibroblasts and Stimulates Monocytes Migration
    Article Snippet: Thermal cycling conditions were: 95 °C for 3 min, followed by 32 cycles of 95 °C for 30 s, 58 °C for 30 s, and 72 °C for 1 min, and followed by a final incubation at 72 °C for 5 min. Amplification products were separated in 1.5% agarose gel in TAE buffer and stained with EthBr, then gel images were captured in a DNR Bio-Imaging System MF-ChemiBIS 2.0. (Mahale HaHamisha, Jerusalem, Israel) using Gel-Capture software. .. In addition, each gene was cloned into a cloning vector pCR2.1® -TOPO® using the TOPO TA Cloning® kit (Invitrogen) and transformed into E. coli Novablue (EMD Chemicals-Novagen Brand., Madison, WI, USA) Then, cloned gene plasmids were purified and used to prepare a standard curve for quantitative RT-PCR (RT-qPCR) in real time.

    Article Title: Equine Arteritis Virus Uses Equine CXCL16 as an Entry Receptor
    Article Snippet: The amplified EqCXCL16 fragment (amino acids [aa] 17 to 247) was run on a 1% agarose gel (E-Gel EX; Life Technologies, Grand Island, NY) and purified with a commercial kit (Zymoclean gel recovery kit; Zymo Research, Irvine, CA). .. The resultant plasmid construct (p15-16A) was used to transform E. coli NovaBlue (EMD Millipore Novagen, Temecula, CA) using a TransformAid kit (Thermo Scientific, Rockford, IL).

    In Vitro:

    Article Title: Integrating artificial with natural cells to translate chemical messages that direct E. coli behaviour
    Article Snippet: Paragraph title: In vitro characterization of the riboswitch ... Plasmids were amplified in E. coli Novablue (Novagen) and purified with Wizard Plus SV Minipreps DNA Purification System (Promega).

    Column Chromatography:

    Article Title: Proteomic Analysis of and Immune Responses to Ehrlichia chaffeensis Lipoproteins
    Article Snippet: The recombinant plasmids were amplified in E. coli NovaBlue (Novagen). .. When the optical density at 600 nm of the E. coli culture reached 0.6, isopropyl-β- d -thiogalactopyranoside (Gold Bio Technology, St. Louis, MO) was added at a final concentration of 1 mM.

    Concentration Assay:

    Article Title: Phospholipase D from Loxosceleslaeta Spider Venom Induces IL-6, IL-8, CXCL1/GRO-α, and CCL2/MCP-1 Production in Human Skin Fibroblasts and Stimulates Monocytes Migration
    Article Snippet: In addition, each gene was cloned into a cloning vector pCR2.1® -TOPO® using the TOPO TA Cloning® kit (Invitrogen) and transformed into E. coli Novablue (EMD Chemicals-Novagen Brand., Madison, WI, USA) Then, cloned gene plasmids were purified and used to prepare a standard curve for quantitative RT-PCR (RT-qPCR) in real time. .. Then, RT-qPCR was then performed using a thermal cycler StepOnePlus™ Real-Time PCR System (Applied Biosystems., Foster City, CA, USA).

    Article Title: Proteomic Analysis of and Immune Responses to Ehrlichia chaffeensis Lipoproteins
    Article Snippet: The recombinant plasmids were amplified in E. coli NovaBlue (Novagen). .. BL21(DE3) cells transformed with lipoprotein constructs were propagated in liquid LB medium in the presence of 50 μg/ml kanamycin (Invitrogen) overnight at 37°C and then subcultured at a 1:200 dilution.

    Article Title: Diversity of hydrolases from hydrothermal vent sediments of the Levante Bay, Vulcano Island (Aeolian archipelago) identified by activity-based metagenomics and biochemical characterization of new esterases and an arabinopyranosidase
    Article Snippet: The purified PCR products were then purified, treated with an endonuclease, annealed to His-tag harbouring the pET-46c Ek/LIC vector, and transformed into E. coli NovaBlue according to the protocol of the manufacturer (Novagen, Darmstadt, Germany). .. The expression of hydrolases in E. coli BL-21 (DE3) was carried out in two steps: at first, the inocula were grown in LB medium supplemented with 100 μg/l ampicillin in an incubator at 37 °C, with shaking at 220 rpm to the OD600 of 0.8–0.9.

    DNA Purification:

    Article Title: Integrating artificial with natural cells to translate chemical messages that direct E. coli behaviour
    Article Snippet: Data are reported as averages with standard error, or representative runs are shown. .. Plasmids were amplified in E. coli Novablue (Novagen) and purified with Wizard Plus SV Minipreps DNA Purification System (Promega). .. Plasmid DNA was phenol–chloroform extracted, ethanol precipitated and resuspended in deionized and diethyl pyrocarbonate-treated water.

    Staining:

    Article Title: Phospholipase D from Loxosceleslaeta Spider Venom Induces IL-6, IL-8, CXCL1/GRO-α, and CCL2/MCP-1 Production in Human Skin Fibroblasts and Stimulates Monocytes Migration
    Article Snippet: Thermal cycling conditions were: 95 °C for 3 min, followed by 32 cycles of 95 °C for 30 s, 58 °C for 30 s, and 72 °C for 1 min, and followed by a final incubation at 72 °C for 5 min. Amplification products were separated in 1.5% agarose gel in TAE buffer and stained with EthBr, then gel images were captured in a DNR Bio-Imaging System MF-ChemiBIS 2.0. (Mahale HaHamisha, Jerusalem, Israel) using Gel-Capture software. .. In addition, each gene was cloned into a cloning vector pCR2.1® -TOPO® using the TOPO TA Cloning® kit (Invitrogen) and transformed into E. coli Novablue (EMD Chemicals-Novagen Brand., Madison, WI, USA) Then, cloned gene plasmids were purified and used to prepare a standard curve for quantitative RT-PCR (RT-qPCR) in real time.

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    Millipore novablue
    Novablue, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 21 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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