nested-pcr strategy Search Results


  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    New England Biolabs pcr based mutagenesis strategy
    Pcr Based Mutagenesis Strategy, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 99/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pcr based mutagenesis strategy/product/New England Biolabs
    Average 99 stars, based on 13 article reviews
    Price from $9.99 to $1999.99
    pcr based mutagenesis strategy - by Bioz Stars, 2020-05
    99/100 stars
      Buy from Supplier

    94
    Thermo Fisher gateway polymerase chain reaction pcr cloning strategy
    Gateway Polymerase Chain Reaction Pcr Cloning Strategy, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/gateway polymerase chain reaction pcr cloning strategy/product/Thermo Fisher
    Average 94 stars, based on 12 article reviews
    Price from $9.99 to $1999.99
    gateway polymerase chain reaction pcr cloning strategy - by Bioz Stars, 2020-05
    94/100 stars
      Buy from Supplier

    88
    PrimerDesign Inc nested pcr strategy
    Nested Pcr Strategy, supplied by PrimerDesign Inc, used in various techniques. Bioz Stars score: 88/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/nested pcr strategy/product/PrimerDesign Inc
    Average 88 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    nested pcr strategy - by Bioz Stars, 2020-05
    88/100 stars
      Buy from Supplier

    86
    Stratagene polymerase chain reaction based mutagenesis strategy
    Polymerase Chain Reaction Based Mutagenesis Strategy, supplied by Stratagene, used in various techniques. Bioz Stars score: 86/100, based on 23 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/polymerase chain reaction based mutagenesis strategy/product/Stratagene
    Average 86 stars, based on 23 article reviews
    Price from $9.99 to $1999.99
    polymerase chain reaction based mutagenesis strategy - by Bioz Stars, 2020-05
    86/100 stars
      Buy from Supplier

    85
    Thermo Fisher fluorescent 5 nuclease real time polymerase chain reaction
    Fluorescent 5 Nuclease Real Time Polymerase Chain Reaction, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/fluorescent 5 nuclease real time polymerase chain reaction/product/Thermo Fisher
    Average 85 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    fluorescent 5 nuclease real time polymerase chain reaction - by Bioz Stars, 2020-05
    85/100 stars
      Buy from Supplier

    85
    Stratagene quickchange pcr strategies
    GILZ is expressed upon stimulation with C3 exotoxin or toxin B. (A) HeLa cells were incubated with toxin B (50ng/ml) for indicated the time spans and immunoblots were performed from whole cell lysates using anti-Rac1 mAb102 recognizing only non-glucosylated Rac-1, and an anti-Rac1 mAb antibody recognizing total Rac1 as well as antibodies recognizing RhoB, actin or GILZ. (B) Immunoblot of GILZ and actin expression upon stimulation of HeLa cells with C2IN-C3lim (100 ng/mL) + C2IIa (200 ng/mL) for indicated time spans. (C) To explore the expression of additional GILZ isoforms, HeLa cells were transfected with 7.5 nM siRNA specific for GILZ or control siRNA for 48 h and subsequently either left untreated or stimulated with C. difficile toxin B (50 ng/ml) or 100 µM DEX for 4 h. Arrows mark three GILZ isoforms which were inhibited by the used GILZ siRNA. Note that only isoform 1 was induced by the used stimuli. (D) Cells were stimulated with toxin B for 2 or 4 h to determine GILZ mRNA expression by real-time <t>RT-PCR.</t> Mean + SD of 2 independent experiments normalized to untreated. (E) To assay transcriptional activity of the GILZ promoter cells were transfected with a luciferase reporter under control of a 2088 bp GILZ promoter and co-transfected with pCMV-ß-gal (for standardization) 24 h before infection with a Y. enterocolitica pYV + and various mutant strains or treatment with DEX or toxin B. Means + SD of 4 independent experiments normalized to untreated. Significant differences compared to untreated are indicated by asterisks (p
    Quickchange Pcr Strategies, supplied by Stratagene, used in various techniques. Bioz Stars score: 85/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/quickchange pcr strategies/product/Stratagene
    Average 85 stars, based on 16 article reviews
    Price from $9.99 to $1999.99
    quickchange pcr strategies - by Bioz Stars, 2020-05
    85/100 stars
      Buy from Supplier

    88
    Stratagene quikchange pcr strategy
    GILZ is expressed upon stimulation with C3 exotoxin or toxin B. (A) HeLa cells were incubated with toxin B (50ng/ml) for indicated the time spans and immunoblots were performed from whole cell lysates using anti-Rac1 mAb102 recognizing only non-glucosylated Rac-1, and an anti-Rac1 mAb antibody recognizing total Rac1 as well as antibodies recognizing RhoB, actin or GILZ. (B) Immunoblot of GILZ and actin expression upon stimulation of HeLa cells with C2IN-C3lim (100 ng/mL) + C2IIa (200 ng/mL) for indicated time spans. (C) To explore the expression of additional GILZ isoforms, HeLa cells were transfected with 7.5 nM siRNA specific for GILZ or control siRNA for 48 h and subsequently either left untreated or stimulated with C. difficile toxin B (50 ng/ml) or 100 µM DEX for 4 h. Arrows mark three GILZ isoforms which were inhibited by the used GILZ siRNA. Note that only isoform 1 was induced by the used stimuli. (D) Cells were stimulated with toxin B for 2 or 4 h to determine GILZ mRNA expression by real-time <t>RT-PCR.</t> Mean + SD of 2 independent experiments normalized to untreated. (E) To assay transcriptional activity of the GILZ promoter cells were transfected with a luciferase reporter under control of a 2088 bp GILZ promoter and co-transfected with pCMV-ß-gal (for standardization) 24 h before infection with a Y. enterocolitica pYV + and various mutant strains or treatment with DEX or toxin B. Means + SD of 4 independent experiments normalized to untreated. Significant differences compared to untreated are indicated by asterisks (p
    Quikchange Pcr Strategy, supplied by Stratagene, used in various techniques. Bioz Stars score: 88/100, based on 39 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/quikchange pcr strategy/product/Stratagene
    Average 88 stars, based on 39 article reviews
    Price from $9.99 to $1999.99
    quikchange pcr strategy - by Bioz Stars, 2020-05
    88/100 stars
      Buy from Supplier

    80
    TaKaRa mimic pcr strategy
    GILZ is expressed upon stimulation with C3 exotoxin or toxin B. (A) HeLa cells were incubated with toxin B (50ng/ml) for indicated the time spans and immunoblots were performed from whole cell lysates using anti-Rac1 mAb102 recognizing only non-glucosylated Rac-1, and an anti-Rac1 mAb antibody recognizing total Rac1 as well as antibodies recognizing RhoB, actin or GILZ. (B) Immunoblot of GILZ and actin expression upon stimulation of HeLa cells with C2IN-C3lim (100 ng/mL) + C2IIa (200 ng/mL) for indicated time spans. (C) To explore the expression of additional GILZ isoforms, HeLa cells were transfected with 7.5 nM siRNA specific for GILZ or control siRNA for 48 h and subsequently either left untreated or stimulated with C. difficile toxin B (50 ng/ml) or 100 µM DEX for 4 h. Arrows mark three GILZ isoforms which were inhibited by the used GILZ siRNA. Note that only isoform 1 was induced by the used stimuli. (D) Cells were stimulated with toxin B for 2 or 4 h to determine GILZ mRNA expression by real-time <t>RT-PCR.</t> Mean + SD of 2 independent experiments normalized to untreated. (E) To assay transcriptional activity of the GILZ promoter cells were transfected with a luciferase reporter under control of a 2088 bp GILZ promoter and co-transfected with pCMV-ß-gal (for standardization) 24 h before infection with a Y. enterocolitica pYV + and various mutant strains or treatment with DEX or toxin B. Means + SD of 4 independent experiments normalized to untreated. Significant differences compared to untreated are indicated by asterisks (p
    Mimic Pcr Strategy, supplied by TaKaRa, used in various techniques. Bioz Stars score: 80/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mimic pcr strategy/product/TaKaRa
    Average 80 stars, based on 7 article reviews
    Price from $9.99 to $1999.99
    mimic pcr strategy - by Bioz Stars, 2020-05
    80/100 stars
      Buy from Supplier

    91
    Stratagene quick change pcr strategy
    GILZ is expressed upon stimulation with C3 exotoxin or toxin B. (A) HeLa cells were incubated with toxin B (50ng/ml) for indicated the time spans and immunoblots were performed from whole cell lysates using anti-Rac1 mAb102 recognizing only non-glucosylated Rac-1, and an anti-Rac1 mAb antibody recognizing total Rac1 as well as antibodies recognizing RhoB, actin or GILZ. (B) Immunoblot of GILZ and actin expression upon stimulation of HeLa cells with C2IN-C3lim (100 ng/mL) + C2IIa (200 ng/mL) for indicated time spans. (C) To explore the expression of additional GILZ isoforms, HeLa cells were transfected with 7.5 nM siRNA specific for GILZ or control siRNA for 48 h and subsequently either left untreated or stimulated with C. difficile toxin B (50 ng/ml) or 100 µM DEX for 4 h. Arrows mark three GILZ isoforms which were inhibited by the used GILZ siRNA. Note that only isoform 1 was induced by the used stimuli. (D) Cells were stimulated with toxin B for 2 or 4 h to determine GILZ mRNA expression by real-time <t>RT-PCR.</t> Mean + SD of 2 independent experiments normalized to untreated. (E) To assay transcriptional activity of the GILZ promoter cells were transfected with a luciferase reporter under control of a 2088 bp GILZ promoter and co-transfected with pCMV-ß-gal (for standardization) 24 h before infection with a Y. enterocolitica pYV + and various mutant strains or treatment with DEX or toxin B. Means + SD of 4 independent experiments normalized to untreated. Significant differences compared to untreated are indicated by asterisks (p
    Quick Change Pcr Strategy, supplied by Stratagene, used in various techniques. Bioz Stars score: 91/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/quick change pcr strategy/product/Stratagene
    Average 91 stars, based on 9 article reviews
    Price from $9.99 to $1999.99
    quick change pcr strategy - by Bioz Stars, 2020-05
    91/100 stars
      Buy from Supplier

    86
    Stratagene pcr based quikchangetm strategy
    GILZ is expressed upon stimulation with C3 exotoxin or toxin B. (A) HeLa cells were incubated with toxin B (50ng/ml) for indicated the time spans and immunoblots were performed from whole cell lysates using anti-Rac1 mAb102 recognizing only non-glucosylated Rac-1, and an anti-Rac1 mAb antibody recognizing total Rac1 as well as antibodies recognizing RhoB, actin or GILZ. (B) Immunoblot of GILZ and actin expression upon stimulation of HeLa cells with C2IN-C3lim (100 ng/mL) + C2IIa (200 ng/mL) for indicated time spans. (C) To explore the expression of additional GILZ isoforms, HeLa cells were transfected with 7.5 nM siRNA specific for GILZ or control siRNA for 48 h and subsequently either left untreated or stimulated with C. difficile toxin B (50 ng/ml) or 100 µM DEX for 4 h. Arrows mark three GILZ isoforms which were inhibited by the used GILZ siRNA. Note that only isoform 1 was induced by the used stimuli. (D) Cells were stimulated with toxin B for 2 or 4 h to determine GILZ mRNA expression by real-time <t>RT-PCR.</t> Mean + SD of 2 independent experiments normalized to untreated. (E) To assay transcriptional activity of the GILZ promoter cells were transfected with a luciferase reporter under control of a 2088 bp GILZ promoter and co-transfected with pCMV-ß-gal (for standardization) 24 h before infection with a Y. enterocolitica pYV + and various mutant strains or treatment with DEX or toxin B. Means + SD of 4 independent experiments normalized to untreated. Significant differences compared to untreated are indicated by asterisks (p
    Pcr Based Quikchangetm Strategy, supplied by Stratagene, used in various techniques. Bioz Stars score: 86/100, based on 29 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pcr based quikchangetm strategy/product/Stratagene
    Average 86 stars, based on 29 article reviews
    Price from $9.99 to $1999.99
    pcr based quikchangetm strategy - by Bioz Stars, 2020-05
    86/100 stars
      Buy from Supplier

    91
    TaKaRa 2 step pcr strategy
    GILZ is expressed upon stimulation with C3 exotoxin or toxin B. (A) HeLa cells were incubated with toxin B (50ng/ml) for indicated the time spans and immunoblots were performed from whole cell lysates using anti-Rac1 mAb102 recognizing only non-glucosylated Rac-1, and an anti-Rac1 mAb antibody recognizing total Rac1 as well as antibodies recognizing RhoB, actin or GILZ. (B) Immunoblot of GILZ and actin expression upon stimulation of HeLa cells with C2IN-C3lim (100 ng/mL) + C2IIa (200 ng/mL) for indicated time spans. (C) To explore the expression of additional GILZ isoforms, HeLa cells were transfected with 7.5 nM siRNA specific for GILZ or control siRNA for 48 h and subsequently either left untreated or stimulated with C. difficile toxin B (50 ng/ml) or 100 µM DEX for 4 h. Arrows mark three GILZ isoforms which were inhibited by the used GILZ siRNA. Note that only isoform 1 was induced by the used stimuli. (D) Cells were stimulated with toxin B for 2 or 4 h to determine GILZ mRNA expression by real-time <t>RT-PCR.</t> Mean + SD of 2 independent experiments normalized to untreated. (E) To assay transcriptional activity of the GILZ promoter cells were transfected with a luciferase reporter under control of a 2088 bp GILZ promoter and co-transfected with pCMV-ß-gal (for standardization) 24 h before infection with a Y. enterocolitica pYV + and various mutant strains or treatment with DEX or toxin B. Means + SD of 4 independent experiments normalized to untreated. Significant differences compared to untreated are indicated by asterisks (p
    2 Step Pcr Strategy, supplied by TaKaRa, used in various techniques. Bioz Stars score: 91/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/2 step pcr strategy/product/TaKaRa
    Average 91 stars, based on 9 article reviews
    Price from $9.99 to $1999.99
    2 step pcr strategy - by Bioz Stars, 2020-05
    91/100 stars
      Buy from Supplier

    95
    Millipore pcr primer strategy
    GILZ is expressed upon stimulation with C3 exotoxin or toxin B. (A) HeLa cells were incubated with toxin B (50ng/ml) for indicated the time spans and immunoblots were performed from whole cell lysates using anti-Rac1 mAb102 recognizing only non-glucosylated Rac-1, and an anti-Rac1 mAb antibody recognizing total Rac1 as well as antibodies recognizing RhoB, actin or GILZ. (B) Immunoblot of GILZ and actin expression upon stimulation of HeLa cells with C2IN-C3lim (100 ng/mL) + C2IIa (200 ng/mL) for indicated time spans. (C) To explore the expression of additional GILZ isoforms, HeLa cells were transfected with 7.5 nM siRNA specific for GILZ or control siRNA for 48 h and subsequently either left untreated or stimulated with C. difficile toxin B (50 ng/ml) or 100 µM DEX for 4 h. Arrows mark three GILZ isoforms which were inhibited by the used GILZ siRNA. Note that only isoform 1 was induced by the used stimuli. (D) Cells were stimulated with toxin B for 2 or 4 h to determine GILZ mRNA expression by real-time <t>RT-PCR.</t> Mean + SD of 2 independent experiments normalized to untreated. (E) To assay transcriptional activity of the GILZ promoter cells were transfected with a luciferase reporter under control of a 2088 bp GILZ promoter and co-transfected with pCMV-ß-gal (for standardization) 24 h before infection with a Y. enterocolitica pYV + and various mutant strains or treatment with DEX or toxin B. Means + SD of 4 independent experiments normalized to untreated. Significant differences compared to untreated are indicated by asterisks (p
    Pcr Primer Strategy, supplied by Millipore, used in various techniques. Bioz Stars score: 95/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pcr primer strategy/product/Millipore
    Average 95 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    pcr primer strategy - by Bioz Stars, 2020-05
    95/100 stars
      Buy from Supplier

    89
    Stratagene pcr strategy
    <t>HIV-1</t> mutant library and NGS sample preparation. (A) Genomic region for each HIV-1 mutation library. (B) The two-step <t>PCR</t> amplicon approach for NGS sample preparation. Virion cDNA from each mutant viral population cell passage is used as template for a HIV-1 specific staggered PCR step that uses primers specific to the HIV-1 mutagenized region containing overhangs with a complex 10 ‘N’ nucleotide tag with two keto “K” or amino “M” nucleotide positions that identify the specific fragment and population, respectively. PCR products from step one are pooled, the amplicon molecule concentration is accurately measured, and then decreased for error correction. The pooled sample is then used as template for a second PCR using a single primer set containing the remainder of the Illumina adapter region for NGS.
    Pcr Strategy, supplied by Stratagene, used in various techniques. Bioz Stars score: 89/100, based on 88 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pcr strategy/product/Stratagene
    Average 89 stars, based on 88 article reviews
    Price from $9.99 to $1999.99
    pcr strategy - by Bioz Stars, 2020-05
    89/100 stars
      Buy from Supplier

    91
    TaKaRa pcr strategy
    Schematic diagram of <t>PTEN</t> localization mutants. To target PTEN into different subcellular compartments, specific localization signals were inserted into PTEN′s sequence via <t>PCR</t> or molecular cloning as described in Materials and Methods. These
    Pcr Strategy, supplied by TaKaRa, used in various techniques. Bioz Stars score: 91/100, based on 29 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pcr strategy/product/TaKaRa
    Average 91 stars, based on 29 article reviews
    Price from $9.99 to $1999.99
    pcr strategy - by Bioz Stars, 2020-05
    91/100 stars
      Buy from Supplier

    89
    The Jackson Laboratory pcr strategy
    Schematic diagram of <t>PTEN</t> localization mutants. To target PTEN into different subcellular compartments, specific localization signals were inserted into PTEN′s sequence via <t>PCR</t> or molecular cloning as described in Materials and Methods. These
    Pcr Strategy, supplied by The Jackson Laboratory, used in various techniques. Bioz Stars score: 89/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pcr strategy/product/The Jackson Laboratory
    Average 89 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    pcr strategy - by Bioz Stars, 2020-05
    89/100 stars
      Buy from Supplier

    88
    Johnson & Johnson pcr strategies
    Schematic diagram of <t>PTEN</t> localization mutants. To target PTEN into different subcellular compartments, specific localization signals were inserted into PTEN′s sequence via <t>PCR</t> or molecular cloning as described in Materials and Methods. These
    Pcr Strategies, supplied by Johnson & Johnson, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pcr strategies/product/Johnson & Johnson
    Average 88 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    pcr strategies - by Bioz Stars, 2020-05
    88/100 stars
      Buy from Supplier

    93
    Addgene inc pcr based strategy
    Schematic diagram of <t>PTEN</t> localization mutants. To target PTEN into different subcellular compartments, specific localization signals were inserted into PTEN′s sequence via <t>PCR</t> or molecular cloning as described in Materials and Methods. These
    Pcr Based Strategy, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pcr based strategy/product/Addgene inc
    Average 93 stars, based on 5 article reviews
    Price from $9.99 to $1999.99
    pcr based strategy - by Bioz Stars, 2020-05
    93/100 stars
      Buy from Supplier

    89
    Thermo Fisher pcr based strategy
    Schematic diagram of <t>PTEN</t> localization mutants. To target PTEN into different subcellular compartments, specific localization signals were inserted into PTEN′s sequence via <t>PCR</t> or molecular cloning as described in Materials and Methods. These
    Pcr Based Strategy, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 89/100, based on 97 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pcr based strategy/product/Thermo Fisher
    Average 89 stars, based on 97 article reviews
    Price from $9.99 to $1999.99
    pcr based strategy - by Bioz Stars, 2020-05
    89/100 stars
      Buy from Supplier

    98
    Thermo Fisher instaclone pcr cloning strategy
    Schematic diagram of <t>PTEN</t> localization mutants. To target PTEN into different subcellular compartments, specific localization signals were inserted into PTEN′s sequence via <t>PCR</t> or molecular cloning as described in Materials and Methods. These
    Instaclone Pcr Cloning Strategy, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 98/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/instaclone pcr cloning strategy/product/Thermo Fisher
    Average 98 stars, based on 6 article reviews
    Price from $9.99 to $1999.99
    instaclone pcr cloning strategy - by Bioz Stars, 2020-05
    98/100 stars
      Buy from Supplier

    Image Search Results


    GILZ is expressed upon stimulation with C3 exotoxin or toxin B. (A) HeLa cells were incubated with toxin B (50ng/ml) for indicated the time spans and immunoblots were performed from whole cell lysates using anti-Rac1 mAb102 recognizing only non-glucosylated Rac-1, and an anti-Rac1 mAb antibody recognizing total Rac1 as well as antibodies recognizing RhoB, actin or GILZ. (B) Immunoblot of GILZ and actin expression upon stimulation of HeLa cells with C2IN-C3lim (100 ng/mL) + C2IIa (200 ng/mL) for indicated time spans. (C) To explore the expression of additional GILZ isoforms, HeLa cells were transfected with 7.5 nM siRNA specific for GILZ or control siRNA for 48 h and subsequently either left untreated or stimulated with C. difficile toxin B (50 ng/ml) or 100 µM DEX for 4 h. Arrows mark three GILZ isoforms which were inhibited by the used GILZ siRNA. Note that only isoform 1 was induced by the used stimuli. (D) Cells were stimulated with toxin B for 2 or 4 h to determine GILZ mRNA expression by real-time RT-PCR. Mean + SD of 2 independent experiments normalized to untreated. (E) To assay transcriptional activity of the GILZ promoter cells were transfected with a luciferase reporter under control of a 2088 bp GILZ promoter and co-transfected with pCMV-ß-gal (for standardization) 24 h before infection with a Y. enterocolitica pYV + and various mutant strains or treatment with DEX or toxin B. Means + SD of 4 independent experiments normalized to untreated. Significant differences compared to untreated are indicated by asterisks (p

    Journal: PLoS ONE

    Article Title: Yersinia enterocolitica YopT and Clostridium difficile Toxin B Induce Expression of GILZ in Epithelial Cells

    doi: 10.1371/journal.pone.0040730

    Figure Lengend Snippet: GILZ is expressed upon stimulation with C3 exotoxin or toxin B. (A) HeLa cells were incubated with toxin B (50ng/ml) for indicated the time spans and immunoblots were performed from whole cell lysates using anti-Rac1 mAb102 recognizing only non-glucosylated Rac-1, and an anti-Rac1 mAb antibody recognizing total Rac1 as well as antibodies recognizing RhoB, actin or GILZ. (B) Immunoblot of GILZ and actin expression upon stimulation of HeLa cells with C2IN-C3lim (100 ng/mL) + C2IIa (200 ng/mL) for indicated time spans. (C) To explore the expression of additional GILZ isoforms, HeLa cells were transfected with 7.5 nM siRNA specific for GILZ or control siRNA for 48 h and subsequently either left untreated or stimulated with C. difficile toxin B (50 ng/ml) or 100 µM DEX for 4 h. Arrows mark three GILZ isoforms which were inhibited by the used GILZ siRNA. Note that only isoform 1 was induced by the used stimuli. (D) Cells were stimulated with toxin B for 2 or 4 h to determine GILZ mRNA expression by real-time RT-PCR. Mean + SD of 2 independent experiments normalized to untreated. (E) To assay transcriptional activity of the GILZ promoter cells were transfected with a luciferase reporter under control of a 2088 bp GILZ promoter and co-transfected with pCMV-ß-gal (for standardization) 24 h before infection with a Y. enterocolitica pYV + and various mutant strains or treatment with DEX or toxin B. Means + SD of 4 independent experiments normalized to untreated. Significant differences compared to untreated are indicated by asterisks (p

    Article Snippet: Point mutations were introduced into the plasmid p1940-Luc by the Quick change PCR based strategy (Stratagene, Cedar Creek, USA) to generate the additional constructs using the following phosphorylated primers: c-myc 1-mut, 5′-GAC GCA GCC GGC TCC TCC-3′, 5′-CCG GGG CGT CAG GGG CCA TGC-3; c-myc 2 mut, 5′GTC CAG GGA GTA TGA CAT GGG AG-3′, 5′-CCG GGC CCT CAC CAT CAC G-3′, Oct-1a, 5′-TGC ATG CCC CTG ACG CTG-3′, 5′-GGC GAG TCC TGT ACC GGG CTT TGT GG-3′; Oct-1b 5′ TGT ATT TCT TAT TTC TCT AGA AAT CAG CTC CAG-3′.

    Techniques: Incubation, Western Blot, Expressing, Transfection, Quantitative RT-PCR, Activity Assay, Luciferase, Infection, Mutagenesis

    The KRAB–KAP-1 repression system targets SETDB1 and enhances H3-K9 methylation and HP1 recruitment to promoters of transcriptionally silenced genes. ( A ) Schematic representation of a two-plasmid system used to create a stably integrated luciferase transgene in NIH/3T3 cells that is regulated by a heterologous KRAB repressor protein. Numbered arrow sets represent the relative position of PCR primers used for PCR amplification of DNA retained by ChIP. ( B ) Two single-cell subclones containing the heterologous KRAB–PAX3–HBD transcriptional repressor and the integrated luciferase transgene, which is either expressed (cl-49) or stably silenced (cl-74) following hormone treatment. Luciferase activities were measured in subconfluent populations of cells and reported as relative light units per milligram of protein. ( C ) ChIP experiments showing the colocalization of KAP-1 and SETDB1 at the TK promoter region of the luciferase transgene in the cells where transcription of the luciferase gene has been stably silenced (cl-74). Formaldehyde cross-linked chromatin from cl-49 and cl-74 cells was immunoprecipitated with either affinity-purified KAP-1 or SETDB1 IgG. An equal amount of promoter sequence in cl-49 and cl-74 nucleosomal preparations was determined by PCR from 1% of the input chromatin. PCR-amplified DNA fragments are illustrated in A . cl-2 represents a negative control cell line. ( D ) ChIPs of cross-linked chromatin with KAP-1, SETDB1, HP1α, and MeK9 antiserum as in C . Bold numbers below each lane represent quantitation of amplified DNA, expressed as percentage of signal intensity for the amplified input DNA.

    Journal: Genes & Development

    Article Title: SETDB1: a novel KAP-1-associated histone H3, lysine 9-specific methyltransferase that contributes to HP1-mediated silencing of euchromatic genes by KRAB zinc-finger proteins

    doi: 10.1101/gad.973302

    Figure Lengend Snippet: The KRAB–KAP-1 repression system targets SETDB1 and enhances H3-K9 methylation and HP1 recruitment to promoters of transcriptionally silenced genes. ( A ) Schematic representation of a two-plasmid system used to create a stably integrated luciferase transgene in NIH/3T3 cells that is regulated by a heterologous KRAB repressor protein. Numbered arrow sets represent the relative position of PCR primers used for PCR amplification of DNA retained by ChIP. ( B ) Two single-cell subclones containing the heterologous KRAB–PAX3–HBD transcriptional repressor and the integrated luciferase transgene, which is either expressed (cl-49) or stably silenced (cl-74) following hormone treatment. Luciferase activities were measured in subconfluent populations of cells and reported as relative light units per milligram of protein. ( C ) ChIP experiments showing the colocalization of KAP-1 and SETDB1 at the TK promoter region of the luciferase transgene in the cells where transcription of the luciferase gene has been stably silenced (cl-74). Formaldehyde cross-linked chromatin from cl-49 and cl-74 cells was immunoprecipitated with either affinity-purified KAP-1 or SETDB1 IgG. An equal amount of promoter sequence in cl-49 and cl-74 nucleosomal preparations was determined by PCR from 1% of the input chromatin. PCR-amplified DNA fragments are illustrated in A . cl-2 represents a negative control cell line. ( D ) ChIPs of cross-linked chromatin with KAP-1, SETDB1, HP1α, and MeK9 antiserum as in C . Bold numbers below each lane represent quantitation of amplified DNA, expressed as percentage of signal intensity for the amplified input DNA.

    Article Snippet: Amino acid substitutions in SETDB1 (R643V, CC 729, 731 LP, H1224K, C1226A, C1279Y) were created using Quick Change PCR mutagenesis strategies (Stratagene).

    Techniques: Methylation, Plasmid Preparation, Stable Transfection, Luciferase, Polymerase Chain Reaction, Amplification, Chromatin Immunoprecipitation, Immunoprecipitation, Affinity Purification, Sequencing, Negative Control, Quantitation Assay

    HIV-1 mutant library and NGS sample preparation. (A) Genomic region for each HIV-1 mutation library. (B) The two-step PCR amplicon approach for NGS sample preparation. Virion cDNA from each mutant viral population cell passage is used as template for a HIV-1 specific staggered PCR step that uses primers specific to the HIV-1 mutagenized region containing overhangs with a complex 10 ‘N’ nucleotide tag with two keto “K” or amino “M” nucleotide positions that identify the specific fragment and population, respectively. PCR products from step one are pooled, the amplicon molecule concentration is accurately measured, and then decreased for error correction. The pooled sample is then used as template for a second PCR using a single primer set containing the remainder of the Illumina adapter region for NGS.

    Journal: Retrovirology

    Article Title: High-throughput profiling of point mutations across the HIV-1 genome

    doi: 10.1186/s12977-014-0124-6

    Figure Lengend Snippet: HIV-1 mutant library and NGS sample preparation. (A) Genomic region for each HIV-1 mutation library. (B) The two-step PCR amplicon approach for NGS sample preparation. Virion cDNA from each mutant viral population cell passage is used as template for a HIV-1 specific staggered PCR step that uses primers specific to the HIV-1 mutagenized region containing overhangs with a complex 10 ‘N’ nucleotide tag with two keto “K” or amino “M” nucleotide positions that identify the specific fragment and population, respectively. PCR products from step one are pooled, the amplicon molecule concentration is accurately measured, and then decreased for error correction. The pooled sample is then used as template for a second PCR using a single primer set containing the remainder of the Illumina adapter region for NGS.

    Article Snippet: Viral mutant library preparation To generate the HIV-1 mutant library we designed a PCR strategy utilizing the HIV-1 proviral DNA plasmid pNL4-3 as template and the error-prone polymerase Mutazyme II (Strategene) to generate the point mutations during PCR amplification.

    Techniques: Mutagenesis, Next-Generation Sequencing, Sample Prep, Polymerase Chain Reaction, Amplification, Concentration Assay

    Changes in PGC-1α expression and acetylation after chronic exercise. A : Real-time PCR analyses of PGC-1 α mRNA expression in the skeletal muscle of C57BL/6J wild-type mice (black columns) and ob / ob mice (gray columns). All data are normalized per 18S rRNA. B : Densitometry of protein data. Homogenates of skeletal muscle were probed with an antibody detecting PGC-1α (90 kDa). Blots were also probed with GAPDH as a loading control. C : Densitometry of immunoprecipitation (IP) experiments performed on skeletal muscle lysates, using PGC-1α for precipitation and an antibody directed against acetyl-lysine or PGC-1α for detection. Data are given as acetylated PGC-1α per total PGC-1α in these samples. All data are means ± SEM. * P

    Journal: Diabetes

    Article Title: Mitochondrial Biogenesis and Peroxisome Proliferator-Activated Receptor-? Coactivator-1? (PGC-1?) Deacetylation by Physical Activity

    doi: 10.2337/db10-0331

    Figure Lengend Snippet: Changes in PGC-1α expression and acetylation after chronic exercise. A : Real-time PCR analyses of PGC-1 α mRNA expression in the skeletal muscle of C57BL/6J wild-type mice (black columns) and ob / ob mice (gray columns). All data are normalized per 18S rRNA. B : Densitometry of protein data. Homogenates of skeletal muscle were probed with an antibody detecting PGC-1α (90 kDa). Blots were also probed with GAPDH as a loading control. C : Densitometry of immunoprecipitation (IP) experiments performed on skeletal muscle lysates, using PGC-1α for precipitation and an antibody directed against acetyl-lysine or PGC-1α for detection. Data are given as acetylated PGC-1α per total PGC-1α in these samples. All data are means ± SEM. * P

    Article Snippet: Site-directed mutations at Thr177 and Ser538 in PGC-1α were introduced by a PCR-based strategy with the QuikChange site-directed mutagenesis kit (Stratagene).

    Techniques: Pyrolysis Gas Chromatography, Expressing, Real-time Polymerase Chain Reaction, Mouse Assay, Immunoprecipitation

    Schematic diagram of PTEN localization mutants. To target PTEN into different subcellular compartments, specific localization signals were inserted into PTEN′s sequence via PCR or molecular cloning as described in Materials and Methods. These

    Journal: Molecular and Cellular Biology

    Article Title: Nuclear PTEN-Mediated Growth Suppression Is Independent of Akt Down-Regulation

    doi: 10.1128/MCB.25.14.6211-6224.2005

    Figure Lengend Snippet: Schematic diagram of PTEN localization mutants. To target PTEN into different subcellular compartments, specific localization signals were inserted into PTEN′s sequence via PCR or molecular cloning as described in Materials and Methods. These

    Article Snippet: The pLNCX-PTEN wild type (WT) and mutants were constructed by initially using a PCR strategy for cloning into the BamHI/EcoRI sites of the pBS (SK+) vector and were subsequently subcloned into the NotI/SalI sites of the pLNCX retroviral vector (Clontech).

    Techniques: Sequencing, Polymerase Chain Reaction, Molecular Cloning