plasmids puc19  (Thermo Fisher)


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    Structured Review

    Thermo Fisher plasmids puc19
    Plasmids Puc19, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 80/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/plasmids puc19/product/Thermo Fisher
    Average 80 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    plasmids puc19 - by Bioz Stars, 2020-01
    80/100 stars

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    Related Articles

    Acetylene Reduction Assay:

    Article Title: Molecular cloning of a novel bioH gene from an environmental metagenome encoding a carboxylesterase with exceptional tolerance to organic solvents
    Article Snippet: Bacterial strains, plasmids and culture conditions Escherichia coli TOP10 [genotype: F¯ mcr A Δ (mrr -hsd RMS-mcr BC) Φ80 lac Z Δ M15 Δ lac X74 rec A1 ara D139 Δ (ara -leu )7697 gal U gal K rps L (StrR ) end A1-nup G] (TianGen Biotech Co. Ltd. Beijing, China) was used as a host strain for transformation of recombinant plasmids in the cloning. .. The plasmid pUC19 (Fermentas, MD, USA) and pET-28a (Merck Millipore, Darmstadt, Germany) were used as cloning and gene expression vector, respectively.

    Amplification:

    Article Title: Quorum-sensing regulation of a capsular polysaccharide receptor for the Rhodobacter capsulatus gene transfer agent (RcGTA)
    Article Snippet: The plasmid pUC19 (Invitrogen) was used for subcloning, pIND4 ( ) as an expression vector for the Δrcc01081 and Δrcc01932 complementation experiments, and pXCA601( ) for the rcc01081 promoter lacZ fusion experiments. .. The insert was made by PCR amplification using WT R. capsulatus genomic DNA as a template, and the primer set 1081_lacZ_for and 1081_lacZ_rev.

    Article Title: Azotobacter vinelandii Nitrogenase Activity, Hydrogen Production, and Response to Oxygen Exposure
    Article Snippet: E. coli TOP10 competent cells (Invitrogen, Carlsbad, CA) were transformed with plasmid pUC19 (Invitrogen) by the heat shock method according to the manufacturer's instructions and plated on LB with ampicillin to select for transformants. .. A region of the A. vinelandii CA6 genome spanning from within phbC to within phbA was amplified by PCR with HindIII and EcoRI sites incorporated into forward and reverse primers, respectively (primers pUCphbCF and pUCphbCR [ ]).

    Article Title: Involvement of Vitamin B6 Biosynthesis Pathways in the Insecticidal Activity of Photorhabdus luminescens
    Article Snippet: The fragment of pdxB with the putative promoter and lambda T0 transcriptional terminator was amplified from pBK-pdxB-T0 using primers pUC19_Pnat_pdxB_InF_F and pUC19_T0_InF_R and ligated into the linearized pUC19 plasmid (linearized by PCR with primers pUC19_linearize_F and pUC19_linearize_R) (Epicentre, WI, USA) by In-Fusion to construct pUC19-pdxB. .. The plasmid pUC19 (blank, without any insertion) or pUC19-pdxB was introduced into the mutant cells by electroporation using EasyjecT Prima (Equibio Ltd., United Kingdom), following the instruction manual for E. coli transformation.

    Clone Assay:

    Article Title: Mdt(A), a New Efflux Protein Conferring Multiple Antibiotic Resistance in Lactococcus lactis and Escherichia coli
    Article Snippet: .. Plasmid pUC19 (Life Technologies) and the shuttle vector pWM401 ( ) were used as cloning vectors. ..

    Article Title: Amylases without known homologues discovered in an acid mine drainage: significance and impact
    Article Snippet: .. The bacteria E. coli DH5α and TOP10 (Invitrogen) and the plasmid pUC19 (Invitrogen) were used as cell host and cloning vector for the subcloning of the putative amylase genes. .. Proteins were expressed in E. coli Arctic DE3 (Stratagene) by using the expressing vector pET30a+.

    Article Title: Molecular cloning of a novel bioH gene from an environmental metagenome encoding a carboxylesterase with exceptional tolerance to organic solvents
    Article Snippet: .. The plasmid pUC19 (Fermentas, MD, USA) and pET-28a (Merck Millipore, Darmstadt, Germany) were used as cloning and gene expression vector, respectively. .. E. coli host cells were grown in Luria-Bertani (LB) liquid medium [ ] at 37°C with shaking at 200 rpm, while E. coli strains containing recombinant plasmids were cultured in LB broth or agar plates supplemented with ampicillin (100 μg/ml) or kanamycin (30 μg/ml).

    Article Title: Involvement of Vitamin B6 Biosynthesis Pathways in the Insecticidal Activity of Photorhabdus luminescens
    Article Snippet: To attach the lambda T0 transcriptional terminator, adaptor-added pdxB gene was ligated into the linearized pBK-miniTn7-T0 with an In-Fusion HD cloning kit (Clontech, CA, USA) to construct pBK-pdxB-T0. .. The plasmid pUC19 (blank, without any insertion) or pUC19-pdxB was introduced into the mutant cells by electroporation using EasyjecT Prima (Equibio Ltd., United Kingdom), following the instruction manual for E. coli transformation.

    Article Title: Secretion of an Endogenous Subtilisin by Pichia pastoris Strains GS115 and KM71 ▿
    Article Snippet: Direct cloning of PCR products in the pDrive vector was performed using a PCR cloning kit (Qiagen, Hombrechtikon, Switzerland). .. All plasmid subcloning experiments were performed with E. coli XL1-Blue with plasmids pUC19 and pPICZA (Invitrogen).

    Construct:

    Article Title: Involvement of Vitamin B6 Biosynthesis Pathways in the Insecticidal Activity of Photorhabdus luminescens
    Article Snippet: The fragment of pdxB with the putative promoter and lambda T0 transcriptional terminator was amplified from pBK-pdxB-T0 using primers pUC19_Pnat_pdxB_InF_F and pUC19_T0_InF_R and ligated into the linearized pUC19 plasmid (linearized by PCR with primers pUC19_linearize_F and pUC19_linearize_R) (Epicentre, WI, USA) by In-Fusion to construct pUC19-pdxB. .. The plasmid pUC19 (blank, without any insertion) or pUC19-pdxB was introduced into the mutant cells by electroporation using EasyjecT Prima (Equibio Ltd., United Kingdom), following the instruction manual for E. coli transformation.

    Article Title: Secretion of an Endogenous Subtilisin by Pichia pastoris Strains GS115 and KM71 ▿
    Article Snippet: Pichia pastoris GS115 and KM71 (Invitrogen, Carlsbad, CA), pPICZA, and previously constructed plasmids pKJ113- LAP2 and pKJ113- DPPIV for expression of the genes encoding Aspergillus fumigatus DppIV (AfuDppIV) and A. fumigatus Lap2 (AfuLap2), respectively, were used ( , ). .. All plasmid subcloning experiments were performed with E. coli XL1-Blue with plasmids pUC19 and pPICZA (Invitrogen).

    Electrophoresis:

    Article Title: Identification of the minimal bacterial 2’-deoxy-7-amido-7-deazaguanine synthesis machinery
    Article Snippet: Purified plasmid pUC19 was linearized by cutting at the single Sal I site using FastDigest Sal I (Thermo) following manufacturer recommended protocols, purified on a 1% agarose gel, and isolated using a GeneJET gel extraction kit (Fermentas). .. The incubations where then subjected to electrophoresis on a 0.5% agarose gel containing 0.33 μg/mL ethidium bromide made with a TBE buffer containing 89 mM Tris (pH 8.3), 89 mM boric acid, and 2 mM EDTA.

    Enzymatic Assay:

    Article Title: Mycofumigation through production of the volatile DNA-methylating agent N-methyl-N-nitrosoisobutyramide by fungi in the genus Muscodor
    Article Snippet: Paragraph title: Enzymatic assay for DNA methylation ... The plasmid pUC19 (Invitrogen) was propagated in E. coli DH5α (Invitrogen), and a Qiagen Plasmid MaxiPrep kit was used to obtain a sufficient amount of plasmid.

    Expressing:

    Article Title: Quorum-sensing regulation of a capsular polysaccharide receptor for the Rhodobacter capsulatus gene transfer agent (RcGTA)
    Article Snippet: .. The plasmid pUC19 (Invitrogen) was used for subcloning, pIND4 ( ) as an expression vector for the Δrcc01081 and Δrcc01932 complementation experiments, and pXCA601( ) for the rcc01081 promoter lacZ fusion experiments. .. The rcc01081::lacZ promoter fusion plasmid containing ~ 1.5 kb of sequence 5′ of the rcc01081 start codon was used to evaluate transcription and translation of the rcc01081 gene.

    Article Title: Amylases without known homologues discovered in an acid mine drainage: significance and impact
    Article Snippet: The bacteria E. coli DH5α and TOP10 (Invitrogen) and the plasmid pUC19 (Invitrogen) were used as cell host and cloning vector for the subcloning of the putative amylase genes. .. Proteins were expressed in E. coli Arctic DE3 (Stratagene) by using the expressing vector pET30a+.

    Article Title: Molecular cloning of a novel bioH gene from an environmental metagenome encoding a carboxylesterase with exceptional tolerance to organic solvents
    Article Snippet: .. The plasmid pUC19 (Fermentas, MD, USA) and pET-28a (Merck Millipore, Darmstadt, Germany) were used as cloning and gene expression vector, respectively. .. E. coli host cells were grown in Luria-Bertani (LB) liquid medium [ ] at 37°C with shaking at 200 rpm, while E. coli strains containing recombinant plasmids were cultured in LB broth or agar plates supplemented with ampicillin (100 μg/ml) or kanamycin (30 μg/ml).

    Article Title: Secretion of an Endogenous Subtilisin by Pichia pastoris Strains GS115 and KM71 ▿
    Article Snippet: Pichia pastoris GS115 and KM71 (Invitrogen, Carlsbad, CA), pPICZA, and previously constructed plasmids pKJ113- LAP2 and pKJ113- DPPIV for expression of the genes encoding Aspergillus fumigatus DppIV (AfuDppIV) and A. fumigatus Lap2 (AfuLap2), respectively, were used ( , ). .. All plasmid subcloning experiments were performed with E. coli XL1-Blue with plasmids pUC19 and pPICZA (Invitrogen).

    Modification:

    Article Title: Quorum-sensing regulation of a capsular polysaccharide receptor for the Rhodobacter capsulatus gene transfer agent (RcGTA)
    Article Snippet: Standard methods of DNA purification, restriction enzyme digestion, and other modification techniques were used ( ). .. The plasmid pUC19 (Invitrogen) was used for subcloning, pIND4 ( ) as an expression vector for the Δrcc01081 and Δrcc01932 complementation experiments, and pXCA601( ) for the rcc01081 promoter lacZ fusion experiments.

    Transformation Assay:

    Article Title: Ectoine can enhance structural changes in DNA in vitro
    Article Snippet: .. Plasmid DNA preparation The plasmid pUC19 (2686 base pairs) was transformed in Escherichia coli Top10 (One Shot® TOP10 Competent Cells, Invitrogen) by using the recommended standard protocol from Invitrogen. .. After cultivating the cells, the plasmids were isolated and purified with NucleoBond® Xtra Midi Plus (Macherey-Nagel) and resuspended in ultra-pure water (conductance 0.055 µS cm−1 , pH 6.6) or in 10 mM Tris buffer (pH 7.5).

    Article Title: Azotobacter vinelandii Nitrogenase Activity, Hydrogen Production, and Response to Oxygen Exposure
    Article Snippet: .. E. coli TOP10 competent cells (Invitrogen, Carlsbad, CA) were transformed with plasmid pUC19 (Invitrogen) by the heat shock method according to the manufacturer's instructions and plated on LB with ampicillin to select for transformants. .. Plasmid was extracted from transformant cultures by the alkaline lysis method using a Qiagen Miniprep kit according to the manufacturer's instructions, and plasmid size was determined by 1% agarose gel electrophoresis with ethidium bromide staining.

    Article Title: Molecular cloning of a novel bioH gene from an environmental metagenome encoding a carboxylesterase with exceptional tolerance to organic solvents
    Article Snippet: Bacterial strains, plasmids and culture conditions Escherichia coli TOP10 [genotype: F¯ mcr A Δ (mrr -hsd RMS-mcr BC) Φ80 lac Z Δ M15 Δ lac X74 rec A1 ara D139 Δ (ara -leu )7697 gal U gal K rps L (StrR ) end A1-nup G] (TianGen Biotech Co. Ltd. Beijing, China) was used as a host strain for transformation of recombinant plasmids in the cloning. .. The plasmid pUC19 (Fermentas, MD, USA) and pET-28a (Merck Millipore, Darmstadt, Germany) were used as cloning and gene expression vector, respectively.

    Article Title: Involvement of Vitamin B6 Biosynthesis Pathways in the Insecticidal Activity of Photorhabdus luminescens
    Article Snippet: .. The plasmid pUC19 (blank, without any insertion) or pUC19-pdxB was introduced into the mutant cells by electroporation using EasyjecT Prima (Equibio Ltd., United Kingdom), following the instruction manual for E. coli transformation. .. Detailed methods for P. luminescens transformation by electroporation are as follows.

    Derivative Assay:

    Article Title: Role of the CipA Scaffoldin Protein in Cellulose Solubilization, as Determined by Targeted Gene Deletion and Complementation in Clostridium thermocellum
    Article Snippet: .. The pMB1 E. coli origin of replication is derived from plasmid pUC19 (Invitrogen Corp.). .. The p15A E. coli origin of replication and arabinose-inducible promoter are derived from plasmid pBAD30 ( ).

    Electroporation:

    Article Title: Involvement of Vitamin B6 Biosynthesis Pathways in the Insecticidal Activity of Photorhabdus luminescens
    Article Snippet: .. The plasmid pUC19 (blank, without any insertion) or pUC19-pdxB was introduced into the mutant cells by electroporation using EasyjecT Prima (Equibio Ltd., United Kingdom), following the instruction manual for E. coli transformation. .. Detailed methods for P. luminescens transformation by electroporation are as follows.

    Cell Culture:

    Article Title: Molecular cloning of a novel bioH gene from an environmental metagenome encoding a carboxylesterase with exceptional tolerance to organic solvents
    Article Snippet: The plasmid pUC19 (Fermentas, MD, USA) and pET-28a (Merck Millipore, Darmstadt, Germany) were used as cloning and gene expression vector, respectively. .. E. coli host cells were grown in Luria-Bertani (LB) liquid medium [ ] at 37°C with shaking at 200 rpm, while E. coli strains containing recombinant plasmids were cultured in LB broth or agar plates supplemented with ampicillin (100 μg/ml) or kanamycin (30 μg/ml).

    Sequencing:

    Article Title: Quorum-sensing regulation of a capsular polysaccharide receptor for the Rhodobacter capsulatus gene transfer agent (RcGTA)
    Article Snippet: The plasmid pUC19 (Invitrogen) was used for subcloning, pIND4 ( ) as an expression vector for the Δrcc01081 and Δrcc01932 complementation experiments, and pXCA601( ) for the rcc01081 promoter lacZ fusion experiments. .. The rcc01081::lacZ promoter fusion plasmid containing ~ 1.5 kb of sequence 5′ of the rcc01081 start codon was used to evaluate transcription and translation of the rcc01081 gene.

    Article Title: Involvement of Vitamin B6 Biosynthesis Pathways in the Insecticidal Activity of Photorhabdus luminescens
    Article Snippet: The coding sequence of the pdxB gene with a putative promoter region ( ) was first amplified by PCR from genomic DNA of wild-type P. luminescens TT01 using primers pdxB_up_F and usg_R and then reamplified from the amplicon using Pnat_pdxB_Inf_F and pdxB_Inf_R to add cloning adaptors. .. The plasmid pUC19 (blank, without any insertion) or pUC19-pdxB was introduced into the mutant cells by electroporation using EasyjecT Prima (Equibio Ltd., United Kingdom), following the instruction manual for E. coli transformation.

    Recombinant:

    Article Title: Quorum-sensing regulation of a capsular polysaccharide receptor for the Rhodobacter capsulatus gene transfer agent (RcGTA)
    Article Snippet: Paragraph title: Recombinant DNA techniques and plasmids ... The plasmid pUC19 (Invitrogen) was used for subcloning, pIND4 ( ) as an expression vector for the Δrcc01081 and Δrcc01932 complementation experiments, and pXCA601( ) for the rcc01081 promoter lacZ fusion experiments.

    Article Title: Molecular cloning of a novel bioH gene from an environmental metagenome encoding a carboxylesterase with exceptional tolerance to organic solvents
    Article Snippet: E. coli BL21(DE3) [genotype: F- omp T hsd SB (r B - m B - ) gal dcm (DE3)] (TianGen) was employed as a host for expression of recombinant proteins. .. The plasmid pUC19 (Fermentas, MD, USA) and pET-28a (Merck Millipore, Darmstadt, Germany) were used as cloning and gene expression vector, respectively.

    Mutagenesis:

    Article Title: Involvement of Vitamin B6 Biosynthesis Pathways in the Insecticidal Activity of Photorhabdus luminescens
    Article Snippet: .. The plasmid pUC19 (blank, without any insertion) or pUC19-pdxB was introduced into the mutant cells by electroporation using EasyjecT Prima (Equibio Ltd., United Kingdom), following the instruction manual for E. coli transformation. .. Detailed methods for P. luminescens transformation by electroporation are as follows.

    Isolation:

    Article Title: Ectoine can enhance structural changes in DNA in vitro
    Article Snippet: Plasmid DNA preparation The plasmid pUC19 (2686 base pairs) was transformed in Escherichia coli Top10 (One Shot® TOP10 Competent Cells, Invitrogen) by using the recommended standard protocol from Invitrogen. .. After cultivating the cells, the plasmids were isolated and purified with NucleoBond® Xtra Midi Plus (Macherey-Nagel) and resuspended in ultra-pure water (conductance 0.055 µS cm−1 , pH 6.6) or in 10 mM Tris buffer (pH 7.5).

    Article Title: Identification of the minimal bacterial 2’-deoxy-7-amido-7-deazaguanine synthesis machinery
    Article Snippet: .. Purified plasmid pUC19 was linearized by cutting at the single Sal I site using FastDigest Sal I (Thermo) following manufacturer recommended protocols, purified on a 1% agarose gel, and isolated using a GeneJET gel extraction kit (Fermentas). .. A portion (0.5 μg) of linearized pUC19 was incubated in a 20 μL reaction at 37 °C for 10 min in the presence of the relevant Dpd protein(s) at a concentration of either 100 nM, 200 nM, 500 nM, 1 μM, 2 μM, or 4 μM, along with 20 mM HEPES (pH 7.5), 50 mM KCl, 10 mM MgCl2 , and 2 mM 2-mercaptoethanol (BME).

    Subcloning:

    Article Title: Quorum-sensing regulation of a capsular polysaccharide receptor for the Rhodobacter capsulatus gene transfer agent (RcGTA)
    Article Snippet: .. The plasmid pUC19 (Invitrogen) was used for subcloning, pIND4 ( ) as an expression vector for the Δrcc01081 and Δrcc01932 complementation experiments, and pXCA601( ) for the rcc01081 promoter lacZ fusion experiments. .. The rcc01081::lacZ promoter fusion plasmid containing ~ 1.5 kb of sequence 5′ of the rcc01081 start codon was used to evaluate transcription and translation of the rcc01081 gene.

    Article Title: Amylases without known homologues discovered in an acid mine drainage: significance and impact
    Article Snippet: .. The bacteria E. coli DH5α and TOP10 (Invitrogen) and the plasmid pUC19 (Invitrogen) were used as cell host and cloning vector for the subcloning of the putative amylase genes. .. Proteins were expressed in E. coli Arctic DE3 (Stratagene) by using the expressing vector pET30a+.

    Article Title: Secretion of an Endogenous Subtilisin by Pichia pastoris Strains GS115 and KM71 ▿
    Article Snippet: .. All plasmid subcloning experiments were performed with E. coli XL1-Blue with plasmids pUC19 and pPICZA (Invitrogen). .. A P. pastoris genomic λEMBL3 library was constructed as previously described for Candida parapsilosis ( ).

    Purification:

    Article Title: Ectoine can enhance structural changes in DNA in vitro
    Article Snippet: Plasmid DNA preparation The plasmid pUC19 (2686 base pairs) was transformed in Escherichia coli Top10 (One Shot® TOP10 Competent Cells, Invitrogen) by using the recommended standard protocol from Invitrogen. .. After cultivating the cells, the plasmids were isolated and purified with NucleoBond® Xtra Midi Plus (Macherey-Nagel) and resuspended in ultra-pure water (conductance 0.055 µS cm−1 , pH 6.6) or in 10 mM Tris buffer (pH 7.5).

    Article Title: Identification of the minimal bacterial 2’-deoxy-7-amido-7-deazaguanine synthesis machinery
    Article Snippet: .. Purified plasmid pUC19 was linearized by cutting at the single Sal I site using FastDigest Sal I (Thermo) following manufacturer recommended protocols, purified on a 1% agarose gel, and isolated using a GeneJET gel extraction kit (Fermentas). .. A portion (0.5 μg) of linearized pUC19 was incubated in a 20 μL reaction at 37 °C for 10 min in the presence of the relevant Dpd protein(s) at a concentration of either 100 nM, 200 nM, 500 nM, 1 μM, 2 μM, or 4 μM, along with 20 mM HEPES (pH 7.5), 50 mM KCl, 10 mM MgCl2 , and 2 mM 2-mercaptoethanol (BME).

    Polymerase Chain Reaction:

    Article Title: Quorum-sensing regulation of a capsular polysaccharide receptor for the Rhodobacter capsulatus gene transfer agent (RcGTA)
    Article Snippet: The plasmid pUC19 (Invitrogen) was used for subcloning, pIND4 ( ) as an expression vector for the Δrcc01081 and Δrcc01932 complementation experiments, and pXCA601( ) for the rcc01081 promoter lacZ fusion experiments. .. The insert was made by PCR amplification using WT R. capsulatus genomic DNA as a template, and the primer set 1081_lacZ_for and 1081_lacZ_rev.

    Article Title: Azotobacter vinelandii Nitrogenase Activity, Hydrogen Production, and Response to Oxygen Exposure
    Article Snippet: E. coli TOP10 competent cells (Invitrogen, Carlsbad, CA) were transformed with plasmid pUC19 (Invitrogen) by the heat shock method according to the manufacturer's instructions and plated on LB with ampicillin to select for transformants. .. A region of the A. vinelandii CA6 genome spanning from within phbC to within phbA was amplified by PCR with HindIII and EcoRI sites incorporated into forward and reverse primers, respectively (primers pUCphbCF and pUCphbCR [ ]).

    Article Title: Involvement of Vitamin B6 Biosynthesis Pathways in the Insecticidal Activity of Photorhabdus luminescens
    Article Snippet: The fragment of pdxB with the putative promoter and lambda T0 transcriptional terminator was amplified from pBK-pdxB-T0 using primers pUC19_Pnat_pdxB_InF_F and pUC19_T0_InF_R and ligated into the linearized pUC19 plasmid (linearized by PCR with primers pUC19_linearize_F and pUC19_linearize_R) (Epicentre, WI, USA) by In-Fusion to construct pUC19-pdxB. .. The plasmid pUC19 (blank, without any insertion) or pUC19-pdxB was introduced into the mutant cells by electroporation using EasyjecT Prima (Equibio Ltd., United Kingdom), following the instruction manual for E. coli transformation.

    Article Title: Secretion of an Endogenous Subtilisin by Pichia pastoris Strains GS115 and KM71 ▿
    Article Snippet: Direct cloning of PCR products in the pDrive vector was performed using a PCR cloning kit (Qiagen, Hombrechtikon, Switzerland). .. All plasmid subcloning experiments were performed with E. coli XL1-Blue with plasmids pUC19 and pPICZA (Invitrogen).

    Article Title: Role of the CipA Scaffoldin Protein in Cellulose Solubilization, as Determined by Targeted Gene Deletion and Complementation in Clostridium thermocellum
    Article Snippet: Sequences of chromosomal DNA were obtained by PCR using genomic DNA from C. thermocellum strain DSM 1313. .. The pMB1 E. coli origin of replication is derived from plasmid pUC19 (Invitrogen Corp.).

    Positron Emission Tomography:

    Article Title: Molecular cloning of a novel bioH gene from an environmental metagenome encoding a carboxylesterase with exceptional tolerance to organic solvents
    Article Snippet: .. The plasmid pUC19 (Fermentas, MD, USA) and pET-28a (Merck Millipore, Darmstadt, Germany) were used as cloning and gene expression vector, respectively. .. E. coli host cells were grown in Luria-Bertani (LB) liquid medium [ ] at 37°C with shaking at 200 rpm, while E. coli strains containing recombinant plasmids were cultured in LB broth or agar plates supplemented with ampicillin (100 μg/ml) or kanamycin (30 μg/ml).

    Staining:

    Article Title: Azotobacter vinelandii Nitrogenase Activity, Hydrogen Production, and Response to Oxygen Exposure
    Article Snippet: E. coli TOP10 competent cells (Invitrogen, Carlsbad, CA) were transformed with plasmid pUC19 (Invitrogen) by the heat shock method according to the manufacturer's instructions and plated on LB with ampicillin to select for transformants. .. Plasmid was extracted from transformant cultures by the alkaline lysis method using a Qiagen Miniprep kit according to the manufacturer's instructions, and plasmid size was determined by 1% agarose gel electrophoresis with ethidium bromide staining.

    Plasmid Preparation:

    Article Title: Quorum-sensing regulation of a capsular polysaccharide receptor for the Rhodobacter capsulatus gene transfer agent (RcGTA)
    Article Snippet: .. The plasmid pUC19 (Invitrogen) was used for subcloning, pIND4 ( ) as an expression vector for the Δrcc01081 and Δrcc01932 complementation experiments, and pXCA601( ) for the rcc01081 promoter lacZ fusion experiments. .. The rcc01081::lacZ promoter fusion plasmid containing ~ 1.5 kb of sequence 5′ of the rcc01081 start codon was used to evaluate transcription and translation of the rcc01081 gene.

    Article Title: Ectoine can enhance structural changes in DNA in vitro
    Article Snippet: .. Plasmid DNA preparation The plasmid pUC19 (2686 base pairs) was transformed in Escherichia coli Top10 (One Shot® TOP10 Competent Cells, Invitrogen) by using the recommended standard protocol from Invitrogen. .. After cultivating the cells, the plasmids were isolated and purified with NucleoBond® Xtra Midi Plus (Macherey-Nagel) and resuspended in ultra-pure water (conductance 0.055 µS cm−1 , pH 6.6) or in 10 mM Tris buffer (pH 7.5).

    Article Title: Mdt(A), a New Efflux Protein Conferring Multiple Antibiotic Resistance in Lactococcus lactis and Escherichia coli
    Article Snippet: .. Plasmid pUC19 (Life Technologies) and the shuttle vector pWM401 ( ) were used as cloning vectors. ..

    Article Title: A Conserved Tyrosyl-Glutamyl Catalytic Dyad in Evolutionarily Linked Enzymes: Carbapenam Synthetase and β-Lactam Synthetase
    Article Snippet: .. Plasmid pUC19 was from Invitrogen (Carlsbad, CA), and pCDFDuet-1 was purchased from Novagen (La Jolla, CA). ..

    Article Title: Amylases without known homologues discovered in an acid mine drainage: significance and impact
    Article Snippet: .. The bacteria E. coli DH5α and TOP10 (Invitrogen) and the plasmid pUC19 (Invitrogen) were used as cell host and cloning vector for the subcloning of the putative amylase genes. .. Proteins were expressed in E. coli Arctic DE3 (Stratagene) by using the expressing vector pET30a+.

    Article Title: Azotobacter vinelandii Nitrogenase Activity, Hydrogen Production, and Response to Oxygen Exposure
    Article Snippet: .. E. coli TOP10 competent cells (Invitrogen, Carlsbad, CA) were transformed with plasmid pUC19 (Invitrogen) by the heat shock method according to the manufacturer's instructions and plated on LB with ampicillin to select for transformants. .. Plasmid was extracted from transformant cultures by the alkaline lysis method using a Qiagen Miniprep kit according to the manufacturer's instructions, and plasmid size was determined by 1% agarose gel electrophoresis with ethidium bromide staining.

    Article Title: OLE RNA protects extremophilic bacteria from alcohol toxicity
    Article Snippet: .. Plasmid pUC19 was purchased from Invitrogen, and pGFPuv was purchased from Clontech. .. Unless otherwise specified, B. halodurans was grown in LB (Lysogeny Broth) broth (USB Corporation) that was prepared at 90% volume, autoclaved and adjusted to full volume and pH ∼10.5 with 10% (w/v) filter-sterilized Na2 CO3 [1% (w/v) final concentration].

    Article Title: Molecular cloning of a novel bioH gene from an environmental metagenome encoding a carboxylesterase with exceptional tolerance to organic solvents
    Article Snippet: .. The plasmid pUC19 (Fermentas, MD, USA) and pET-28a (Merck Millipore, Darmstadt, Germany) were used as cloning and gene expression vector, respectively. .. E. coli host cells were grown in Luria-Bertani (LB) liquid medium [ ] at 37°C with shaking at 200 rpm, while E. coli strains containing recombinant plasmids were cultured in LB broth or agar plates supplemented with ampicillin (100 μg/ml) or kanamycin (30 μg/ml).

    Article Title: Detection of Ampicillin-Resistant E. coli Using Novel Nanoprobe-Combined Fluorescence In Situ Hybridization
    Article Snippet: .. The E. coli DH5α and plasmid pUC19 were purchased from Invitrogen (Carlsbad, CA, USA, Cat. no. 18258-012). .. Cell Fixation The bacterial cell fixation was performed by modified fixation method [ ].

    Article Title: Mycofumigation through production of the volatile DNA-methylating agent N-methyl-N-nitrosoisobutyramide by fungi in the genus Muscodor
    Article Snippet: .. The plasmid pUC19 (Invitrogen) was propagated in E. coli DH5α (Invitrogen), and a Qiagen Plasmid MaxiPrep kit was used to obtain a sufficient amount of plasmid. .. 2 ml of 25 ng/μl plasmid pUC19 in 10 m m HEPES, pH 7.5, 50 m m NaCl were incubated on one side of a split vial system and exposed to volatiles from a 3-day-old Muscodor isolate (P912B, M. albus , or P1813B) or 1.5-ml microcentrifuge tube caps containing varying amounts of MNIBA (0.01 to 1 mg) in pump oil with a final volume of 100 μl.

    Article Title: Identification of the minimal bacterial 2’-deoxy-7-amido-7-deazaguanine synthesis machinery
    Article Snippet: .. Purified plasmid pUC19 was linearized by cutting at the single Sal I site using FastDigest Sal I (Thermo) following manufacturer recommended protocols, purified on a 1% agarose gel, and isolated using a GeneJET gel extraction kit (Fermentas). .. A portion (0.5 μg) of linearized pUC19 was incubated in a 20 μL reaction at 37 °C for 10 min in the presence of the relevant Dpd protein(s) at a concentration of either 100 nM, 200 nM, 500 nM, 1 μM, 2 μM, or 4 μM, along with 20 mM HEPES (pH 7.5), 50 mM KCl, 10 mM MgCl2 , and 2 mM 2-mercaptoethanol (BME).

    Article Title: Characterization of the dsDNA prophage sequences in the genome of Neisseria gonorrhoeae and visualization of productive bacteriophage
    Article Snippet: .. Plasmid pUC19 was purchased from MBI Fermentas. .. Plasmid pXYL20 was described previously [ ].

    Article Title: Involvement of Vitamin B6 Biosynthesis Pathways in the Insecticidal Activity of Photorhabdus luminescens
    Article Snippet: .. The plasmid pUC19 (blank, without any insertion) or pUC19-pdxB was introduced into the mutant cells by electroporation using EasyjecT Prima (Equibio Ltd., United Kingdom), following the instruction manual for E. coli transformation. .. Detailed methods for P. luminescens transformation by electroporation are as follows.

    Article Title: Secretion of an Endogenous Subtilisin by Pichia pastoris Strains GS115 and KM71 ▿
    Article Snippet: .. All plasmid subcloning experiments were performed with E. coli XL1-Blue with plasmids pUC19 and pPICZA (Invitrogen). .. A P. pastoris genomic λEMBL3 library was constructed as previously described for Candida parapsilosis ( ).

    Article Title: Role of the CipA Scaffoldin Protein in Cellulose Solubilization, as Determined by Targeted Gene Deletion and Complementation in Clostridium thermocellum
    Article Snippet: .. The pMB1 E. coli origin of replication is derived from plasmid pUC19 (Invitrogen Corp.). .. The p15A E. coli origin of replication and arabinose-inducible promoter are derived from plasmid pBAD30 ( ).

    Binding Assay:

    Article Title: Identification of the minimal bacterial 2’-deoxy-7-amido-7-deazaguanine synthesis machinery
    Article Snippet: Paragraph title: DNA binding experiments ... Purified plasmid pUC19 was linearized by cutting at the single Sal I site using FastDigest Sal I (Thermo) following manufacturer recommended protocols, purified on a 1% agarose gel, and isolated using a GeneJET gel extraction kit (Fermentas).

    Agarose Gel Electrophoresis:

    Article Title: Azotobacter vinelandii Nitrogenase Activity, Hydrogen Production, and Response to Oxygen Exposure
    Article Snippet: E. coli TOP10 competent cells (Invitrogen, Carlsbad, CA) were transformed with plasmid pUC19 (Invitrogen) by the heat shock method according to the manufacturer's instructions and plated on LB with ampicillin to select for transformants. .. Plasmid was extracted from transformant cultures by the alkaline lysis method using a Qiagen Miniprep kit according to the manufacturer's instructions, and plasmid size was determined by 1% agarose gel electrophoresis with ethidium bromide staining.

    Article Title: Identification of the minimal bacterial 2’-deoxy-7-amido-7-deazaguanine synthesis machinery
    Article Snippet: .. Purified plasmid pUC19 was linearized by cutting at the single Sal I site using FastDigest Sal I (Thermo) following manufacturer recommended protocols, purified on a 1% agarose gel, and isolated using a GeneJET gel extraction kit (Fermentas). .. A portion (0.5 μg) of linearized pUC19 was incubated in a 20 μL reaction at 37 °C for 10 min in the presence of the relevant Dpd protein(s) at a concentration of either 100 nM, 200 nM, 500 nM, 1 μM, 2 μM, or 4 μM, along with 20 mM HEPES (pH 7.5), 50 mM KCl, 10 mM MgCl2 , and 2 mM 2-mercaptoethanol (BME).

    Incubation:

    Article Title: Mycofumigation through production of the volatile DNA-methylating agent N-methyl-N-nitrosoisobutyramide by fungi in the genus Muscodor
    Article Snippet: The plasmid pUC19 (Invitrogen) was propagated in E. coli DH5α (Invitrogen), and a Qiagen Plasmid MaxiPrep kit was used to obtain a sufficient amount of plasmid. .. 2 ml of 25 ng/μl plasmid pUC19 in 10 m m HEPES, pH 7.5, 50 m m NaCl were incubated on one side of a split vial system and exposed to volatiles from a 3-day-old Muscodor isolate (P912B, M. albus , or P1813B) or 1.5-ml microcentrifuge tube caps containing varying amounts of MNIBA (0.01 to 1 mg) in pump oil with a final volume of 100 μl.

    Article Title: Identification of the minimal bacterial 2’-deoxy-7-amido-7-deazaguanine synthesis machinery
    Article Snippet: Purified plasmid pUC19 was linearized by cutting at the single Sal I site using FastDigest Sal I (Thermo) following manufacturer recommended protocols, purified on a 1% agarose gel, and isolated using a GeneJET gel extraction kit (Fermentas). .. A portion (0.5 μg) of linearized pUC19 was incubated in a 20 μL reaction at 37 °C for 10 min in the presence of the relevant Dpd protein(s) at a concentration of either 100 nM, 200 nM, 500 nM, 1 μM, 2 μM, or 4 μM, along with 20 mM HEPES (pH 7.5), 50 mM KCl, 10 mM MgCl2 , and 2 mM 2-mercaptoethanol (BME).

    DNA Methylation Assay:

    Article Title: Mycofumigation through production of the volatile DNA-methylating agent N-methyl-N-nitrosoisobutyramide by fungi in the genus Muscodor
    Article Snippet: Paragraph title: Enzymatic assay for DNA methylation ... The plasmid pUC19 (Invitrogen) was propagated in E. coli DH5α (Invitrogen), and a Qiagen Plasmid MaxiPrep kit was used to obtain a sufficient amount of plasmid.

    Concentration Assay:

    Article Title: Ectoine can enhance structural changes in DNA in vitro
    Article Snippet: Plasmid DNA preparation The plasmid pUC19 (2686 base pairs) was transformed in Escherichia coli Top10 (One Shot® TOP10 Competent Cells, Invitrogen) by using the recommended standard protocol from Invitrogen. .. The concentration and purity of plasmids was determined by using NanoDrop2000c (Thermo Scientific).

    Article Title: OLE RNA protects extremophilic bacteria from alcohol toxicity
    Article Snippet: Plasmid pUC19 was purchased from Invitrogen, and pGFPuv was purchased from Clontech. .. Unless otherwise specified, B. halodurans was grown in LB (Lysogeny Broth) broth (USB Corporation) that was prepared at 90% volume, autoclaved and adjusted to full volume and pH ∼10.5 with 10% (w/v) filter-sterilized Na2 CO3 [1% (w/v) final concentration].

    Article Title: Identification of the minimal bacterial 2’-deoxy-7-amido-7-deazaguanine synthesis machinery
    Article Snippet: Purified plasmid pUC19 was linearized by cutting at the single Sal I site using FastDigest Sal I (Thermo) following manufacturer recommended protocols, purified on a 1% agarose gel, and isolated using a GeneJET gel extraction kit (Fermentas). .. A portion (0.5 μg) of linearized pUC19 was incubated in a 20 μL reaction at 37 °C for 10 min in the presence of the relevant Dpd protein(s) at a concentration of either 100 nM, 200 nM, 500 nM, 1 μM, 2 μM, or 4 μM, along with 20 mM HEPES (pH 7.5), 50 mM KCl, 10 mM MgCl2 , and 2 mM 2-mercaptoethanol (BME).

    Alkaline Lysis:

    Article Title: Azotobacter vinelandii Nitrogenase Activity, Hydrogen Production, and Response to Oxygen Exposure
    Article Snippet: E. coli TOP10 competent cells (Invitrogen, Carlsbad, CA) were transformed with plasmid pUC19 (Invitrogen) by the heat shock method according to the manufacturer's instructions and plated on LB with ampicillin to select for transformants. .. Plasmid was extracted from transformant cultures by the alkaline lysis method using a Qiagen Miniprep kit according to the manufacturer's instructions, and plasmid size was determined by 1% agarose gel electrophoresis with ethidium bromide staining.

    DNA Purification:

    Article Title: Quorum-sensing regulation of a capsular polysaccharide receptor for the Rhodobacter capsulatus gene transfer agent (RcGTA)
    Article Snippet: Standard methods of DNA purification, restriction enzyme digestion, and other modification techniques were used ( ). .. The plasmid pUC19 (Invitrogen) was used for subcloning, pIND4 ( ) as an expression vector for the Δrcc01081 and Δrcc01932 complementation experiments, and pXCA601( ) for the rcc01081 promoter lacZ fusion experiments.

    Gel Extraction:

    Article Title: Identification of the minimal bacterial 2’-deoxy-7-amido-7-deazaguanine synthesis machinery
    Article Snippet: .. Purified plasmid pUC19 was linearized by cutting at the single Sal I site using FastDigest Sal I (Thermo) following manufacturer recommended protocols, purified on a 1% agarose gel, and isolated using a GeneJET gel extraction kit (Fermentas). .. A portion (0.5 μg) of linearized pUC19 was incubated in a 20 μL reaction at 37 °C for 10 min in the presence of the relevant Dpd protein(s) at a concentration of either 100 nM, 200 nM, 500 nM, 1 μM, 2 μM, or 4 μM, along with 20 mM HEPES (pH 7.5), 50 mM KCl, 10 mM MgCl2 , and 2 mM 2-mercaptoethanol (BME).

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    Thermo Fisher puc19 dna
    Results from <t>DNA</t> damage assay and abiotic ROS generation (A) The plasmid <t>pUC19</t> was treated with the different Cu species for 24 hours and the resulting DNA species were separated and analyzed using gel electrophoresis. UV and restriction enzyme ( Pst I) were used as positive controls for fragmented and linearized DNA, respectively. Electrophoresis was run at 5 V/cm for 1 hour. N-Cu and m-Cu generated the most severe DNA damage and resulted in complete degradation of the plasmid DNA. N-Cu(OH) 2 -a and n-Cu(OH) 2 -b induced complete conversion of supercoiled DNA to open circular and linear DNA, while exposure to the other Cu species resulted in only partial conversion of the plasmid DNA. (B) The capability of each particle to generate reactive oxygen species (ROS) was tested in vitro using (2’,7’ – dichlorofluorescein) DCFH. The particles were treated with DCFH for 2 hours and the fluorescence intensity, which reflects the extent of oxidation, was measured using excitation/emission wavelengths of 530/630 nm. Three replicates were performed at each concentration of Cu species.
    Puc19 Dna, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/puc19 dna/product/Thermo Fisher
    Average 90 stars, based on 6 article reviews
    Price from $9.99 to $1999.99
    puc19 dna - by Bioz Stars, 2020-01
    90/100 stars
      Buy from Supplier

    90
    Thermo Fisher puc19 plasmid dna
    Results from <t>DNA</t> damage assay and abiotic ROS generation (A) The plasmid <t>pUC19</t> was treated with the different Cu species for 24 hours and the resulting DNA species were separated and analyzed using gel electrophoresis. UV and restriction enzyme ( Pst I) were used as positive controls for fragmented and linearized DNA, respectively. Electrophoresis was run at 5 V/cm for 1 hour. N-Cu and m-Cu generated the most severe DNA damage and resulted in complete degradation of the plasmid DNA. N-Cu(OH) 2 -a and n-Cu(OH) 2 -b induced complete conversion of supercoiled DNA to open circular and linear DNA, while exposure to the other Cu species resulted in only partial conversion of the plasmid DNA. (B) The capability of each particle to generate reactive oxygen species (ROS) was tested in vitro using (2’,7’ – dichlorofluorescein) DCFH. The particles were treated with DCFH for 2 hours and the fluorescence intensity, which reflects the extent of oxidation, was measured using excitation/emission wavelengths of 530/630 nm. Three replicates were performed at each concentration of Cu species.
    Puc19 Plasmid Dna, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 22 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/puc19 plasmid dna/product/Thermo Fisher
    Average 90 stars, based on 22 article reviews
    Price from $9.99 to $1999.99
    puc19 plasmid dna - by Bioz Stars, 2020-01
    90/100 stars
      Buy from Supplier

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    Results from DNA damage assay and abiotic ROS generation (A) The plasmid pUC19 was treated with the different Cu species for 24 hours and the resulting DNA species were separated and analyzed using gel electrophoresis. UV and restriction enzyme ( Pst I) were used as positive controls for fragmented and linearized DNA, respectively. Electrophoresis was run at 5 V/cm for 1 hour. N-Cu and m-Cu generated the most severe DNA damage and resulted in complete degradation of the plasmid DNA. N-Cu(OH) 2 -a and n-Cu(OH) 2 -b induced complete conversion of supercoiled DNA to open circular and linear DNA, while exposure to the other Cu species resulted in only partial conversion of the plasmid DNA. (B) The capability of each particle to generate reactive oxygen species (ROS) was tested in vitro using (2’,7’ – dichlorofluorescein) DCFH. The particles were treated with DCFH for 2 hours and the fluorescence intensity, which reflects the extent of oxidation, was measured using excitation/emission wavelengths of 530/630 nm. Three replicates were performed at each concentration of Cu species.

    Journal: ACS nano

    Article Title: Cu Nanoparticles Have Different Impacts in E. coli and L. brevis than Their Micron-Sized and Ionic Analogs

    doi: 10.1021/acsnano.5b02021

    Figure Lengend Snippet: Results from DNA damage assay and abiotic ROS generation (A) The plasmid pUC19 was treated with the different Cu species for 24 hours and the resulting DNA species were separated and analyzed using gel electrophoresis. UV and restriction enzyme ( Pst I) were used as positive controls for fragmented and linearized DNA, respectively. Electrophoresis was run at 5 V/cm for 1 hour. N-Cu and m-Cu generated the most severe DNA damage and resulted in complete degradation of the plasmid DNA. N-Cu(OH) 2 -a and n-Cu(OH) 2 -b induced complete conversion of supercoiled DNA to open circular and linear DNA, while exposure to the other Cu species resulted in only partial conversion of the plasmid DNA. (B) The capability of each particle to generate reactive oxygen species (ROS) was tested in vitro using (2’,7’ – dichlorofluorescein) DCFH. The particles were treated with DCFH for 2 hours and the fluorescence intensity, which reflects the extent of oxidation, was measured using excitation/emission wavelengths of 530/630 nm. Three replicates were performed at each concentration of Cu species.

    Article Snippet: Purified plasmid pUC19 (Thermo Scientific, catalog #SD0061) was incubated in the presence of 100 mg/L Cu species for 24 hours in purified water.

    Techniques: Plasmid Preparation, Nucleic Acid Electrophoresis, Electrophoresis, Generated, In Vitro, Fluorescence, Concentration Assay