one-step rt-pcr kit Search Results


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  • 99
    Thermo Fisher superscript one step rt pcr kit
    p53-mediated induction of Apaf1 mRNA in neurons. (A) <t>RNA</t> was extracted from neurons 36 or 48 h after infection with Ad-p53 or Ad-p53-173L and analyzed for Apaf1 or GAPDH expression using semiquantitative <t>RT-PCR.</t> (B) RNA was extracted from wild-type or p53-deficient neurons at the indicated times after treatment with 10 μM camptothecin and analyzed for Apaf1 or GAPDH expression using semiquantitative RT-PCR.
    Superscript One Step Rt Pcr Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 2504 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Qiagen one step rt pcr kit
    Nested <t>PCR</t> for <t>WT1</t> expression.
    One Step Rt Pcr Kit, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 12955 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Bio-Rad iscript one step rt pcr kit
    In utero B(a)P-exposure upregulates heart nNOS, eNOS, BH4/BH2 oxidore-ductase and AngII mRNA in offspring. Relative expression levels of nNOS, eNOS, BH4/BH2 and AngII as measured by <t>qRT-PCR.</t> Quantitative RT-PCR was performed using <t>iScript</t> one-step RT-PCR
    Iscript One Step Rt Pcr Kit, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 94/100, based on 5922 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher agpath id one step rt pcr kit
    Effect of enzyme system on detection of pathogen targets in primary clinical specimens. Data shown are difference in Ct value between reactions using Quanta One-step <t>RT-PCR</t> ToughMix and <t>AgPath-ID</t> One-step RT-PCR kit when testing TNA extracted from NP/OP swabs (A) or blood (B). Each data point represents the difference in Ct value between the two reactions for an individual clinical specimen. Median difference is indicated ( ― ) for assays with ≥2 positive results. *Targets that were only detected using AgPath always occurred when Ct values were > 33.
    Agpath Id One Step Rt Pcr Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1859 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher superscript iii one step rt pcr kit
    Full-length AKAP13 mRNA levels are reduced by the gene-trap events. (A) TaqMan gene expression assays were used to measure the expression of AKAP13 transcripts at the indicated exon-exon junctions (E4-5, Brx-9, E37-38). (B) Quantitative <t>PCR</t> analysis of wild-type (WT), heterozygote (Het) and homozygote (Hom) neonatal mouse heart and lung RNA for AKAP13 showed that none of the gene-trap mutations affected expression of the E4-5 exon-exon junction. The ΔBrx gene-trap dose dependently decreased expression of the Brx-9 exon-exon junction. Expression of the Brx-9 junction was eliminated in the AKAP13 ΔBrx/ΔBrx mice. All <t>three</t> gene-traps decreased expression of the E37-38 exon-exon junction in a dose-dependent manner. Expression of the E37-38 junction was eliminated in the AKAP13 ΔGEF/ΔGEF mice. The means and standard deviations are graphed for six mice per genotype. One-way ANOVA and Bonferroni’s multiple comparison tests were conducted (Prism 5; GraphPad). †, p
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    Thermo Fisher one step rt pcr kit
    Expression of TANK1 and TANK1a in WT and TANK1 −/− mouse tissues. All <t>RNA</t> samples were serially diluted as indicated for <t>PCR</t> amplification and analysis. Actin cDNA was used as an RT-PCR loading control in all experiments. (A) RT-PCR analysis for TANK1 mRNA expression in various tissues of WT and TANK1 −/− (KO) mice as indicated. 5′-TANK1 RT-PCR products represent upstream cDNA of TANK1. (B) RT-PCR analysis for TANK1 mRNA expression in WT and TANK1 −/− (KO) testis as indicated. 5′-TANK1 and 3′-TANK1 RT-PCR products represent upstream and downstream cDNAs of TANK1; and TANK1a RT-PCR products are specific for TANK1a. (C) RT-PCR analysis for TANK1a mRNA expression in various tissues of WT mice as indicated. TANK1a RT-PCR products represent cDNA of TANK1a. (D, E) Western blot analysis was used to determine tankyrase 1 and 1a expression in thymus, testis and spleen of WT and TANK1 −/− (KO) mice with 762 (anti-SAM, D) and 376 (anti-HPS, E) antibodies. TANK1 indicates tankyrase 1 protein, TANK1x indicates a possible degraded tankyrase 1 protein, and TANK1a indicates tankyrase 1a protein.
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    TaKaRa one step sybr primescript rt pcr kit
    Melting peaks of the <t>SYBR</t> Green Ⅰ-based one-step <t>qRT-PCR</t> products. ( A ) Melting peaks of the qRT-PCR products obtained from in vitro transcribed HTNV cRNA ranging from 1×10 8 to 1×10 3 copies/µl. Only a single sharp peak at 84°C was visible in the melt peak chart. ( B ) Melting peaks obtained from serum samples of HFRS patients. All melting curves in A and B have similar shapes.
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    TaKaRa one step rt pcr kit
    PGRN deficiency increased the production of systemic and local pro‐inflammatory mediators after injection of LPS . ELISA of serum levels of inflammatory mediators including tumour necrosis factor α ( TNF ‐α) ( A ), interleukin 6 ( IL ‐6) ( B ), IL ‐10 ( C ) and monocyte chemoattractant protein 1 ( MCP ‐1) ( D ). ( E ) Western blot analysis of cold‐inducible <t>RNA</t> ‐binding protein ( CIRP ) and high mobility group box protein 1 ( HMGB 1) in the serum from WT and Grn −/− mice with LPS injection at 6 and 24 hrs. PS red, Ponceau S red staining. Real‐time RT ‐ <t>PCR</t> analysis of mRNA levels of TNF ‐α ( F ), IL ‐6 ( G ), IL ‐1β ( H ) and IL ‐10 ( I ) in the lung after LPS injection. Data are mean ± S.D. * P
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    Thermo Fisher vetmax plus one step rt pcr kit
    PGRN deficiency increased the production of systemic and local pro‐inflammatory mediators after injection of LPS . ELISA of serum levels of inflammatory mediators including tumour necrosis factor α ( TNF ‐α) ( A ), interleukin 6 ( IL ‐6) ( B ), IL ‐10 ( C ) and monocyte chemoattractant protein 1 ( MCP ‐1) ( D ). ( E ) Western blot analysis of cold‐inducible <t>RNA</t> ‐binding protein ( CIRP ) and high mobility group box protein 1 ( HMGB 1) in the serum from WT and Grn −/− mice with LPS injection at 6 and 24 hrs. PS red, Ponceau S red staining. Real‐time RT ‐ <t>PCR</t> analysis of mRNA levels of TNF ‐α ( F ), IL ‐6 ( G ), IL ‐1β ( H ) and IL ‐10 ( I ) in the lung after LPS injection. Data are mean ± S.D. * P
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    TaKaRa primescript one step rt pcr kit
    PGRN deficiency increased the production of systemic and local pro‐inflammatory mediators after injection of LPS . ELISA of serum levels of inflammatory mediators including tumour necrosis factor α ( TNF ‐α) ( A ), interleukin 6 ( IL ‐6) ( B ), IL ‐10 ( C ) and monocyte chemoattractant protein 1 ( MCP ‐1) ( D ). ( E ) Western blot analysis of cold‐inducible <t>RNA</t> ‐binding protein ( CIRP ) and high mobility group box protein 1 ( HMGB 1) in the serum from WT and Grn −/− mice with LPS injection at 6 and 24 hrs. PS red, Ponceau S red staining. Real‐time RT ‐ <t>PCR</t> analysis of mRNA levels of TNF ‐α ( F ), IL ‐6 ( G ), IL ‐1β ( H ) and IL ‐10 ( I ) in the lung after LPS injection. Data are mean ± S.D. * P
    Primescript One Step Rt Pcr Kit, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 1089 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher taqman one step rt pcr master mix reagents kit
    Overexpression of dominant negative NRF-1 attenuates the increase in β1 subunit mRNA levels in response to neuronal stimulation of primary cortical neurons. Primary cortical neurons were transfected with empty vector (pcDNA3) or dominant negative NRF-1 (NRF-1 DN; using Nucleofection™) and plated in 6-well plates. DIV7 cells were treated with either vehicle or 20 mM KCl for 6 h. Total mRNA was isolated from cells and quantified by <t>TaqMan</t> <t>qRT-PCR.</t> Gabrb1 mRNA expression was normalized to Cyclophilin A mRNA levels and is presented relative to its levels in pcDNA3 transfected neurons that were treated with vehicle (expressed as 1). Data represent the average ± SEM of n = 3 independent primary neuronal cultures. Two-way ANOVA was performed (comparison between treatments, p = 0.0148; comparison between transfected DNA conditions, p = 0.0031) with Bonferroni post hoc analysis (adjusted p value = 0.0263 for KCl vs. water with pcDNA transfection, not significant for NRF-1 DN).
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    Thermo Fisher taqman one step rt pcr kit
    Overexpression of dominant negative NRF-1 attenuates the increase in β1 subunit mRNA levels in response to neuronal stimulation of primary cortical neurons. Primary cortical neurons were transfected with empty vector (pcDNA3) or dominant negative NRF-1 (NRF-1 DN; using Nucleofection™) and plated in 6-well plates. DIV7 cells were treated with either vehicle or 20 mM KCl for 6 h. Total mRNA was isolated from cells and quantified by <t>TaqMan</t> <t>qRT-PCR.</t> Gabrb1 mRNA expression was normalized to Cyclophilin A mRNA levels and is presented relative to its levels in pcDNA3 transfected neurons that were treated with vehicle (expressed as 1). Data represent the average ± SEM of n = 3 independent primary neuronal cultures. Two-way ANOVA was performed (comparison between treatments, p = 0.0148; comparison between transfected DNA conditions, p = 0.0031) with Bonferroni post hoc analysis (adjusted p value = 0.0263 for KCl vs. water with pcDNA transfection, not significant for NRF-1 DN).
    Taqman One Step Rt Pcr Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 436 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Qiagen one step quantitect sybr green rt pcr kit
    NGF or BDNF changes the gene expression profile of BMSC and increases IL-6 mRNA. BMSC were treated with 100 ng/ml of (A) NGF or (B) BDNF for 18 hours. The cells were collected, RNA was isolated, and microarray analyses were performed. Graphs summarize the panel of gene expression changes in the BMSC treated with NGF or BDNF compared to untreated control cells. BMSC were treated with 100 ng/ml of (C) NGF or (D) BDNF for 2, 4 and 8 hours. The cells were collected, RNA was isolated and real time <t>PCR</t> was performed with the one-step QuantiTech <t>SYBR</t> Green kit as instructed by the manufacturer. BMSC IL-6 expression in response to NGF or BDNF exposure compared to untreated control cells is shown. Expression was normalized to the housekeeping gene GUSB . Statistical analysis was completed by ANOVA ( P ≤0.0001) with significance indicated by an asterisk.
    One Step Quantitect Sybr Green Rt Pcr Kit, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 469 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher path id multiplex one step rt pcr kit
    Timing of non‐biologic product–associated cases of Theiler's disease and predicted exposure periods. Ten non‐biologic‐associated cases of Theiler's disease were diagnosed on 6 properties (properties A‐F) between January 2014 and February 2018. Each property is indicated by a different color. On 4 properties, there was follow‐up monitoring of in‐contact horses by examination, serum chemistry, and equine parvovirus‐hepatitis <t>quantitative‐PCR.</t> The time span during which new cases of hepatitis were identified on each farm is shown above the timeline (diamonds indicate that only 1 case occurred on that farm). In equine biologic product–associated cases, hepatitis typically occurs 4‐10 weeks after administration of an equine biologic product. Assuming a similar time frame between exposure and disease for non‐biologic‐associated cases, a predicted timeline of exposure for each farm was constructed, calculated as 10 weeks before the 1st case to 4 weeks before the last case
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    Bio-Rad one step rt pcr kit
    HIV-2 Vpx represses TLR-mediated IL6, IL12p40, and TNFα induction. THP-1 cells infected for 48 h with equal amounts of HIV-2 viruses with or without vpx and pseudotyped with the VSVg envelope ( A and B ) were treated with TLR ligands for various times as indicated, and then total RNA was isolated, and mRNA levels were measured by <t>qRT-PCR</t> of IL6, IL12p40, and TNFα ( A ) or FLIP or IκBα, and β-actin (as an internal control) ( B ). Each mRNA induction was calculated based on untreated THP-1 cells. Each RT-PCR was run in triplicate. Data are shown as one representative example of three independent experiments, and values are mean ± S.D. C, total DNA was isolated from HIV-2-infected cells, and the level of viral DNA was determined by measuring Gag gene using real time qPCR.
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    Bio-Rad iscripttm one step rt pcr kit
    HIV-2 Vpx represses TLR-mediated IL6, IL12p40, and TNFα induction. THP-1 cells infected for 48 h with equal amounts of HIV-2 viruses with or without vpx and pseudotyped with the VSVg envelope ( A and B ) were treated with TLR ligands for various times as indicated, and then total RNA was isolated, and mRNA levels were measured by <t>qRT-PCR</t> of IL6, IL12p40, and TNFα ( A ) or FLIP or IκBα, and β-actin (as an internal control) ( B ). Each mRNA induction was calculated based on untreated THP-1 cells. Each RT-PCR was run in triplicate. Data are shown as one representative example of three independent experiments, and values are mean ± S.D. C, total DNA was isolated from HIV-2-infected cells, and the level of viral DNA was determined by measuring Gag gene using real time qPCR.
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    TaKaRa titanium one step rt pcr kit
    Relative expression of PKD isoforms in <t>HeLa</t> cells and their depletion by siRNA. (A) Analysis of mRNA expression by <t>RT-PCR</t> shows that PKD2 and PKD3 are the only PKD isoforms expressed in HeLa cells. RT-PCR reaction without the reverse transcriptase (RT−) was used as a negative control and PCR with the corresponding PKD cDNA (C) as a positive control. (B) Quantitative real-time RT-PCR analysis was performed on RNA extracted from HeLa cells. Bars represent the mean (±SD) of the relative mRNA expression of each PKD isoform compared with the average expression of β-actin. *, P
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    Meridian Life Science mytaq one step rt pcr kit
    Relative expression of PKD isoforms in <t>HeLa</t> cells and their depletion by siRNA. (A) Analysis of mRNA expression by <t>RT-PCR</t> shows that PKD2 and PKD3 are the only PKD isoforms expressed in HeLa cells. RT-PCR reaction without the reverse transcriptase (RT−) was used as a negative control and PCR with the corresponding PKD cDNA (C) as a positive control. (B) Quantitative real-time RT-PCR analysis was performed on RNA extracted from HeLa cells. Bars represent the mean (±SD) of the relative mRNA expression of each PKD isoform compared with the average expression of β-actin. *, P
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    Thermo Fisher superscript iii platinum one step quantitative rt pcr kit
    Sensitivity analysis of <t>three</t> reverse primers used in the <t>rRT-PCR</t> assay. Cq values are shown against a ten-fold serial dilution of OROV RNA, either from the prototype strain, or an Ecuadorian strain.
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    Image Search Results


    p53-mediated induction of Apaf1 mRNA in neurons. (A) RNA was extracted from neurons 36 or 48 h after infection with Ad-p53 or Ad-p53-173L and analyzed for Apaf1 or GAPDH expression using semiquantitative RT-PCR. (B) RNA was extracted from wild-type or p53-deficient neurons at the indicated times after treatment with 10 μM camptothecin and analyzed for Apaf1 or GAPDH expression using semiquantitative RT-PCR.

    Journal: The Journal of Cell Biology

    Article Title: APAF1 is a key transcriptional target for p53 in the regulation of neuronal cell death

    doi: 10.1083/jcb.200105137

    Figure Lengend Snippet: p53-mediated induction of Apaf1 mRNA in neurons. (A) RNA was extracted from neurons 36 or 48 h after infection with Ad-p53 or Ad-p53-173L and analyzed for Apaf1 or GAPDH expression using semiquantitative RT-PCR. (B) RNA was extracted from wild-type or p53-deficient neurons at the indicated times after treatment with 10 μM camptothecin and analyzed for Apaf1 or GAPDH expression using semiquantitative RT-PCR.

    Article Snippet: 2 ng of total RNA was used for cDNA synthesis and targeted gene amplification using the SuperScript One-Step RT-PCR kit (GIBCO BRL). cDNA synthesis was carried out at 48°C for 45 min followed by a 2 min initial denaturation step at 94°C.

    Techniques: Infection, Expressing, Reverse Transcription Polymerase Chain Reaction

    Nested PCR for WT1 expression.

    Journal: Biomarker Insights

    Article Title: Studies of Wilms' Tumor (WT1) Gene Expression in Adult Acute Leukemias in Singapore

    doi:

    Figure Lengend Snippet: Nested PCR for WT1 expression.

    Article Snippet: Nested RT-PCR (reverse-transcription PCR) One step nested reverse transcription polymerase chain reaction (Nested RT-PCR) was performed by using a one-step RT-PCR kit. (Qiagen, Valencia, CA, USA) WT1 exon 1–4 was amplified using forward (5′-CCTACCTGCCC AG CTGCCTC-3′) and reverse (5′-CTCCTAAGTTCATCTGATTCC-3′) primers for 20 cycles (annealing temperature 56 °C), followed by nested PCR (forward: 5′-AGAGCCAGCCCGCTATTCG-3′; and reverse: GGTCATGCATTCAAGCTGG-3′ primers) for 30 cycles (annealing temperature 58 °C).

    Techniques: Nested PCR, Expressing

    In utero B(a)P-exposure upregulates heart nNOS, eNOS, BH4/BH2 oxidore-ductase and AngII mRNA in offspring. Relative expression levels of nNOS, eNOS, BH4/BH2 and AngII as measured by qRT-PCR. Quantitative RT-PCR was performed using iScript one-step RT-PCR

    Journal: Toxicology

    Article Title: In utero exposure to benzo(a)pyrene predisposes offspring to cardiovascular dysfunction in later-life

    doi: 10.1016/j.tox.2012.01.017

    Figure Lengend Snippet: In utero B(a)P-exposure upregulates heart nNOS, eNOS, BH4/BH2 oxidore-ductase and AngII mRNA in offspring. Relative expression levels of nNOS, eNOS, BH4/BH2 and AngII as measured by qRT-PCR. Quantitative RT-PCR was performed using iScript one-step RT-PCR

    Article Snippet: Quantitative Real time RT-PCR was carried out using the iScript one-step RT-PCR kit with SYBR Green, PCR plates and optically clear adhesive plate seals (Bio-Rad Laboratories, USA).

    Techniques: In Utero, Expressing, Quantitative RT-PCR, Reverse Transcription Polymerase Chain Reaction

    Effect of enzyme system on detection of pathogen targets in primary clinical specimens. Data shown are difference in Ct value between reactions using Quanta One-step RT-PCR ToughMix and AgPath-ID One-step RT-PCR kit when testing TNA extracted from NP/OP swabs (A) or blood (B). Each data point represents the difference in Ct value between the two reactions for an individual clinical specimen. Median difference is indicated ( ― ) for assays with ≥2 positive results. *Targets that were only detected using AgPath always occurred when Ct values were > 33.

    Journal: PLoS ONE

    Article Title: Optimization of Multiple Pathogen Detection Using the TaqMan Array Card: Application for a Population-Based Study of Neonatal Infection

    doi: 10.1371/journal.pone.0066183

    Figure Lengend Snippet: Effect of enzyme system on detection of pathogen targets in primary clinical specimens. Data shown are difference in Ct value between reactions using Quanta One-step RT-PCR ToughMix and AgPath-ID One-step RT-PCR kit when testing TNA extracted from NP/OP swabs (A) or blood (B). Each data point represents the difference in Ct value between the two reactions for an individual clinical specimen. Median difference is indicated ( ― ) for assays with ≥2 positive results. *Targets that were only detected using AgPath always occurred when Ct values were > 33.

    Article Snippet: We compared the performance of two enzyme formulations, Ambion AgPath-ID One-step RT-PCR kit (Applied Biosystems) and Quanta qScript XLT One-step RT-qPCR ToughMix, low ROX (Quanta Biosciences), at detecting targets in NP/OP specimens (n = 18) and blood specimens (n = 12) ( ).

    Techniques: Reverse Transcription Polymerase Chain Reaction

    Full-length AKAP13 mRNA levels are reduced by the gene-trap events. (A) TaqMan gene expression assays were used to measure the expression of AKAP13 transcripts at the indicated exon-exon junctions (E4-5, Brx-9, E37-38). (B) Quantitative PCR analysis of wild-type (WT), heterozygote (Het) and homozygote (Hom) neonatal mouse heart and lung RNA for AKAP13 showed that none of the gene-trap mutations affected expression of the E4-5 exon-exon junction. The ΔBrx gene-trap dose dependently decreased expression of the Brx-9 exon-exon junction. Expression of the Brx-9 junction was eliminated in the AKAP13 ΔBrx/ΔBrx mice. All three gene-traps decreased expression of the E37-38 exon-exon junction in a dose-dependent manner. Expression of the E37-38 junction was eliminated in the AKAP13 ΔGEF/ΔGEF mice. The means and standard deviations are graphed for six mice per genotype. One-way ANOVA and Bonferroni’s multiple comparison tests were conducted (Prism 5; GraphPad). †, p

    Journal: PLoS ONE

    Article Title: AKAP13 Rho-GEF and PKD-Binding Domain Deficient Mice Develop Normally but Have an Abnormal Response to ?-Adrenergic-Induced Cardiac Hypertrophy

    doi: 10.1371/journal.pone.0062705

    Figure Lengend Snippet: Full-length AKAP13 mRNA levels are reduced by the gene-trap events. (A) TaqMan gene expression assays were used to measure the expression of AKAP13 transcripts at the indicated exon-exon junctions (E4-5, Brx-9, E37-38). (B) Quantitative PCR analysis of wild-type (WT), heterozygote (Het) and homozygote (Hom) neonatal mouse heart and lung RNA for AKAP13 showed that none of the gene-trap mutations affected expression of the E4-5 exon-exon junction. The ΔBrx gene-trap dose dependently decreased expression of the Brx-9 exon-exon junction. Expression of the Brx-9 junction was eliminated in the AKAP13 ΔBrx/ΔBrx mice. All three gene-traps decreased expression of the E37-38 exon-exon junction in a dose-dependent manner. Expression of the E37-38 junction was eliminated in the AKAP13 ΔGEF/ΔGEF mice. The means and standard deviations are graphed for six mice per genotype. One-way ANOVA and Bonferroni’s multiple comparison tests were conducted (Prism 5; GraphPad). †, p

    Article Snippet: Total RNA was extracted from ES cells with Trizol (Invitrogen), and RT-PCR was conducted using the SuperScript III One-Step RT-PCR kit (Invitrogen).

    Techniques: Expressing, Real-time Polymerase Chain Reaction, Mouse Assay

    Gene-traps disrupt AKAP13 in multiple locations. (A) Schematic of the AKAP13 genomic locus. Exons are depicted with black bars, cassette exons with a grey box, and alternative promoters with arrows. The three gene-trap insertions are indicated. (B) Diagram of the gene-trap constructs (blue boxes) integrated between AKAP13 exons (open boxes with exon numbers). The gene-trap vector contains a strong splice acceptor (SA), βGeo cassette (β−galactosidase and neomyocin resistance genes), and stop codon, as well as a polyadenylation (pA) sequence. The splicing events indicated were confirmed by RT-PCR and sequencing. Primers used to genotype the wild-type and gene-trap alleles are shown (black arrows). (C) Resulting protein fusions of AKAP-Lbc, Brx, and Lbc isoforms with βGeo for the gene-trap mutational series. PKA = protein kinase A binding domain, GEF = Rho-guanine nucleotide exchange factor domain, PKD = protein kinase D binding domain, LZ = leucine zipper domain. (D) Sample genotyping of mouse tail clips for the AKAP13 gene-trap mutations using primers in (B). WT = Wild-type, Het = Heterozygote, Hom = Homozygote.

    Journal: PLoS ONE

    Article Title: AKAP13 Rho-GEF and PKD-Binding Domain Deficient Mice Develop Normally but Have an Abnormal Response to ?-Adrenergic-Induced Cardiac Hypertrophy

    doi: 10.1371/journal.pone.0062705

    Figure Lengend Snippet: Gene-traps disrupt AKAP13 in multiple locations. (A) Schematic of the AKAP13 genomic locus. Exons are depicted with black bars, cassette exons with a grey box, and alternative promoters with arrows. The three gene-trap insertions are indicated. (B) Diagram of the gene-trap constructs (blue boxes) integrated between AKAP13 exons (open boxes with exon numbers). The gene-trap vector contains a strong splice acceptor (SA), βGeo cassette (β−galactosidase and neomyocin resistance genes), and stop codon, as well as a polyadenylation (pA) sequence. The splicing events indicated were confirmed by RT-PCR and sequencing. Primers used to genotype the wild-type and gene-trap alleles are shown (black arrows). (C) Resulting protein fusions of AKAP-Lbc, Brx, and Lbc isoforms with βGeo for the gene-trap mutational series. PKA = protein kinase A binding domain, GEF = Rho-guanine nucleotide exchange factor domain, PKD = protein kinase D binding domain, LZ = leucine zipper domain. (D) Sample genotyping of mouse tail clips for the AKAP13 gene-trap mutations using primers in (B). WT = Wild-type, Het = Heterozygote, Hom = Homozygote.

    Article Snippet: Total RNA was extracted from ES cells with Trizol (Invitrogen), and RT-PCR was conducted using the SuperScript III One-Step RT-PCR kit (Invitrogen).

    Techniques: Construct, Plasmid Preparation, Sequencing, Reverse Transcription Polymerase Chain Reaction, Binding Assay

    Expression of TANK1 and TANK1a in WT and TANK1 −/− mouse tissues. All RNA samples were serially diluted as indicated for PCR amplification and analysis. Actin cDNA was used as an RT-PCR loading control in all experiments. (A) RT-PCR analysis for TANK1 mRNA expression in various tissues of WT and TANK1 −/− (KO) mice as indicated. 5′-TANK1 RT-PCR products represent upstream cDNA of TANK1. (B) RT-PCR analysis for TANK1 mRNA expression in WT and TANK1 −/− (KO) testis as indicated. 5′-TANK1 and 3′-TANK1 RT-PCR products represent upstream and downstream cDNAs of TANK1; and TANK1a RT-PCR products are specific for TANK1a. (C) RT-PCR analysis for TANK1a mRNA expression in various tissues of WT mice as indicated. TANK1a RT-PCR products represent cDNA of TANK1a. (D, E) Western blot analysis was used to determine tankyrase 1 and 1a expression in thymus, testis and spleen of WT and TANK1 −/− (KO) mice with 762 (anti-SAM, D) and 376 (anti-HPS, E) antibodies. TANK1 indicates tankyrase 1 protein, TANK1x indicates a possible degraded tankyrase 1 protein, and TANK1a indicates tankyrase 1a protein.

    Journal: PLoS ONE

    Article Title: Tankyrase 1 and Tankyrase 2 Are Essential but Redundant for Mouse Embryonic Development

    doi: 10.1371/journal.pone.0002639

    Figure Lengend Snippet: Expression of TANK1 and TANK1a in WT and TANK1 −/− mouse tissues. All RNA samples were serially diluted as indicated for PCR amplification and analysis. Actin cDNA was used as an RT-PCR loading control in all experiments. (A) RT-PCR analysis for TANK1 mRNA expression in various tissues of WT and TANK1 −/− (KO) mice as indicated. 5′-TANK1 RT-PCR products represent upstream cDNA of TANK1. (B) RT-PCR analysis for TANK1 mRNA expression in WT and TANK1 −/− (KO) testis as indicated. 5′-TANK1 and 3′-TANK1 RT-PCR products represent upstream and downstream cDNAs of TANK1; and TANK1a RT-PCR products are specific for TANK1a. (C) RT-PCR analysis for TANK1a mRNA expression in various tissues of WT mice as indicated. TANK1a RT-PCR products represent cDNA of TANK1a. (D, E) Western blot analysis was used to determine tankyrase 1 and 1a expression in thymus, testis and spleen of WT and TANK1 −/− (KO) mice with 762 (anti-SAM, D) and 376 (anti-HPS, E) antibodies. TANK1 indicates tankyrase 1 protein, TANK1x indicates a possible degraded tankyrase 1 protein, and TANK1a indicates tankyrase 1a protein.

    Article Snippet: RT-PCR reactions were carried out in 50 µl of PCR reaction mix containing 25 µl of PCR buffer (One-step RT-PCR kit, Life Technologies), 1 µg of total RNA and 0.2 µM primers.

    Techniques: Expressing, Polymerase Chain Reaction, Amplification, Reverse Transcription Polymerase Chain Reaction, Mouse Assay, Western Blot

    Melting peaks of the SYBR Green Ⅰ-based one-step qRT-PCR products. ( A ) Melting peaks of the qRT-PCR products obtained from in vitro transcribed HTNV cRNA ranging from 1×10 8 to 1×10 3 copies/µl. Only a single sharp peak at 84°C was visible in the melt peak chart. ( B ) Melting peaks obtained from serum samples of HFRS patients. All melting curves in A and B have similar shapes.

    Journal: PLoS ONE

    Article Title: Quantification of Hantaan Virus with a SYBR Green I-Based One-Step qRT-PCR Assay

    doi: 10.1371/journal.pone.0081525

    Figure Lengend Snippet: Melting peaks of the SYBR Green Ⅰ-based one-step qRT-PCR products. ( A ) Melting peaks of the qRT-PCR products obtained from in vitro transcribed HTNV cRNA ranging from 1×10 8 to 1×10 3 copies/µl. Only a single sharp peak at 84°C was visible in the melt peak chart. ( B ) Melting peaks obtained from serum samples of HFRS patients. All melting curves in A and B have similar shapes.

    Article Snippet: SYBR Green Ⅰ-based one-step qRT-PCR assay qRT-PCR amplification was performed using a Bio-Rad iQ5 Multi-color Real-Time PCR Detection System (Bio-Rad, Hercules, CA, USA) with the One-Step SYBR PrimeScript RT-PCR Kit ii (TaKaRa Biotechnology, Dalian, China).

    Techniques: SYBR Green Assay, Quantitative RT-PCR, In Vitro

    Sensitivity of the SYBR Green Ⅰ-based one-step qRT-PCR assay. ( A ) Serial dilutions of the HTNV cRNA standards ranging from 1×10 6 to 1×10 -1 copies/µl were amplified using the SYBR Green Ⅰ-based one-step qRT-PCR assay in duplicate, and the minimum detection limit of the qRT-PCR assay was 10 copies/µl. ( B ) The qRT-PCR products were electrophoresed on a 2% agarose gel. Marker: DNA ladder of DL2000.

    Journal: PLoS ONE

    Article Title: Quantification of Hantaan Virus with a SYBR Green I-Based One-Step qRT-PCR Assay

    doi: 10.1371/journal.pone.0081525

    Figure Lengend Snippet: Sensitivity of the SYBR Green Ⅰ-based one-step qRT-PCR assay. ( A ) Serial dilutions of the HTNV cRNA standards ranging from 1×10 6 to 1×10 -1 copies/µl were amplified using the SYBR Green Ⅰ-based one-step qRT-PCR assay in duplicate, and the minimum detection limit of the qRT-PCR assay was 10 copies/µl. ( B ) The qRT-PCR products were electrophoresed on a 2% agarose gel. Marker: DNA ladder of DL2000.

    Article Snippet: SYBR Green Ⅰ-based one-step qRT-PCR assay qRT-PCR amplification was performed using a Bio-Rad iQ5 Multi-color Real-Time PCR Detection System (Bio-Rad, Hercules, CA, USA) with the One-Step SYBR PrimeScript RT-PCR Kit ii (TaKaRa Biotechnology, Dalian, China).

    Techniques: SYBR Green Assay, Quantitative RT-PCR, Amplification, Agarose Gel Electrophoresis, Marker

    Amplification chart and melt peak chart of HTNV and SEOV. ( A ) Different dilutions of HTNV and SEOV RNA were tested using the SYBR Green Ⅰ-based one-step qRT-PCR assay, and only HTNV RNA was amplified. ( B ) A single sharp peak at 84°C in the melt peak chart indicated that only specific products of HTNV was obtained.

    Journal: PLoS ONE

    Article Title: Quantification of Hantaan Virus with a SYBR Green I-Based One-Step qRT-PCR Assay

    doi: 10.1371/journal.pone.0081525

    Figure Lengend Snippet: Amplification chart and melt peak chart of HTNV and SEOV. ( A ) Different dilutions of HTNV and SEOV RNA were tested using the SYBR Green Ⅰ-based one-step qRT-PCR assay, and only HTNV RNA was amplified. ( B ) A single sharp peak at 84°C in the melt peak chart indicated that only specific products of HTNV was obtained.

    Article Snippet: SYBR Green Ⅰ-based one-step qRT-PCR assay qRT-PCR amplification was performed using a Bio-Rad iQ5 Multi-color Real-Time PCR Detection System (Bio-Rad, Hercules, CA, USA) with the One-Step SYBR PrimeScript RT-PCR Kit ii (TaKaRa Biotechnology, Dalian, China).

    Techniques: Amplification, SYBR Green Assay, Quantitative RT-PCR

    Expression profiles of ALDH3 family members in human tissues. cDNAs from human liver, kidney, and brain tissue (human MTC (multiple tissue cDNA) panels, Takara Bio) ( A ) or prepared from primary keratinocytes (undifferentiated or differentiated for 7 days; CELLnTEC) ( B ) were subjected to SYBR Green-based real-time quantitative PCR using specific primers for ALDH3A1 , ALDH3A2 , ALDH3B1 , and GAPDH . Values represent the means ± S.D. from three independent reactions. A statistically significant difference is indicated (**, p

    Journal: The Journal of Biological Chemistry

    Article Title: Disruption of the Sjögren-Larsson Syndrome Gene Aldh3a2 in Mice Increases Keratinocyte Growth and Retards Skin Barrier Recovery *

    doi: 10.1074/jbc.M116.714030

    Figure Lengend Snippet: Expression profiles of ALDH3 family members in human tissues. cDNAs from human liver, kidney, and brain tissue (human MTC (multiple tissue cDNA) panels, Takara Bio) ( A ) or prepared from primary keratinocytes (undifferentiated or differentiated for 7 days; CELLnTEC) ( B ) were subjected to SYBR Green-based real-time quantitative PCR using specific primers for ALDH3A1 , ALDH3A2 , ALDH3B1 , and GAPDH . Values represent the means ± S.D. from three independent reactions. A statistically significant difference is indicated (**, p

    Article Snippet: Real-time quantitative PCR was performed using the One-Step SYBR PrimeScript RT-PCR Kit II (Takara Bio) on a CFX96 Touch real-time PCR detection system (Bio-Rad) according to the manufacturer's manual.

    Techniques: Expressing, SYBR Green Assay, Real-time Polymerase Chain Reaction

    Enhanced proliferation and oxidative stress response in Aldh3a2 −/− keratinocytes. A , primary keratinocytes were prepared from wild-type and Aldh3a2 KO mice and grown. Cell numbers were counted by microscopic observation. For each dish, cell numbers in three randomly chosen viewing fields were counted and summed. Values represent the means ± S.D. from four independent experiments. B , keratinocytes prepared from wild-type and Aldh3a2 KO mice were subjected to a [ 3 H]thymidine uptake assay. Values represent the means ± S.D. from three independent experiments. C and D , total RNAs prepared from wild-type and Aldh3a2 KO keratinocytes were subjected to SYBR Green-based real-time quantitative RT-PCR using specific primers for Ki67 ( C ), Hmox1 , Sod1 , Gclc , Gclm , Gsta1 , and Gapdh ( D ). Values are the amount of each mRNA relative to that of Gapdh and represent the means ± S.D. for three independent experiments. Statistically significant differences are indicated (**, p

    Journal: The Journal of Biological Chemistry

    Article Title: Disruption of the Sjögren-Larsson Syndrome Gene Aldh3a2 in Mice Increases Keratinocyte Growth and Retards Skin Barrier Recovery *

    doi: 10.1074/jbc.M116.714030

    Figure Lengend Snippet: Enhanced proliferation and oxidative stress response in Aldh3a2 −/− keratinocytes. A , primary keratinocytes were prepared from wild-type and Aldh3a2 KO mice and grown. Cell numbers were counted by microscopic observation. For each dish, cell numbers in three randomly chosen viewing fields were counted and summed. Values represent the means ± S.D. from four independent experiments. B , keratinocytes prepared from wild-type and Aldh3a2 KO mice were subjected to a [ 3 H]thymidine uptake assay. Values represent the means ± S.D. from three independent experiments. C and D , total RNAs prepared from wild-type and Aldh3a2 KO keratinocytes were subjected to SYBR Green-based real-time quantitative RT-PCR using specific primers for Ki67 ( C ), Hmox1 , Sod1 , Gclc , Gclm , Gsta1 , and Gapdh ( D ). Values are the amount of each mRNA relative to that of Gapdh and represent the means ± S.D. for three independent experiments. Statistically significant differences are indicated (**, p

    Article Snippet: Real-time quantitative PCR was performed using the One-Step SYBR PrimeScript RT-PCR Kit II (Takara Bio) on a CFX96 Touch real-time PCR detection system (Bio-Rad) according to the manufacturer's manual.

    Techniques: Mouse Assay, SYBR Green Assay, Quantitative RT-PCR

    Expression profiles of Aldh3 family members in various tissues. Total RNAs prepared from liver, kidney, retina, cornea, brain, small intestine, lung, testis, spleen, dermis, and epidermis tissue ( A and C ) and from keratinocytes kept undifferentiated or differentiated for 4 days ( B and D ) obtained from wild-type ( A–D ) and Aldh3a2 KO mice ( C and D ) were subjected to SYBR Green-based real-time quantitative RT-PCR using specific primers for Aldh3a1 , Aldh3a2 , Aldh3b1 , Aldh3b2 , Aldh3b3 , and Gapdh . Values are the amount of each mRNA relative to that of Gapdh , and represent the means ± S.D. for three independent experiments. A statistically significant difference is indicated (**, p

    Journal: The Journal of Biological Chemistry

    Article Title: Disruption of the Sjögren-Larsson Syndrome Gene Aldh3a2 in Mice Increases Keratinocyte Growth and Retards Skin Barrier Recovery *

    doi: 10.1074/jbc.M116.714030

    Figure Lengend Snippet: Expression profiles of Aldh3 family members in various tissues. Total RNAs prepared from liver, kidney, retina, cornea, brain, small intestine, lung, testis, spleen, dermis, and epidermis tissue ( A and C ) and from keratinocytes kept undifferentiated or differentiated for 4 days ( B and D ) obtained from wild-type ( A–D ) and Aldh3a2 KO mice ( C and D ) were subjected to SYBR Green-based real-time quantitative RT-PCR using specific primers for Aldh3a1 , Aldh3a2 , Aldh3b1 , Aldh3b2 , Aldh3b3 , and Gapdh . Values are the amount of each mRNA relative to that of Gapdh , and represent the means ± S.D. for three independent experiments. A statistically significant difference is indicated (**, p

    Article Snippet: Real-time quantitative PCR was performed using the One-Step SYBR PrimeScript RT-PCR Kit II (Takara Bio) on a CFX96 Touch real-time PCR detection system (Bio-Rad) according to the manufacturer's manual.

    Techniques: Expressing, Mouse Assay, SYBR Green Assay, Quantitative RT-PCR

    PGRN deficiency increased the production of systemic and local pro‐inflammatory mediators after injection of LPS . ELISA of serum levels of inflammatory mediators including tumour necrosis factor α ( TNF ‐α) ( A ), interleukin 6 ( IL ‐6) ( B ), IL ‐10 ( C ) and monocyte chemoattractant protein 1 ( MCP ‐1) ( D ). ( E ) Western blot analysis of cold‐inducible RNA ‐binding protein ( CIRP ) and high mobility group box protein 1 ( HMGB 1) in the serum from WT and Grn −/− mice with LPS injection at 6 and 24 hrs. PS red, Ponceau S red staining. Real‐time RT ‐ PCR analysis of mRNA levels of TNF ‐α ( F ), IL ‐6 ( G ), IL ‐1β ( H ) and IL ‐10 ( I ) in the lung after LPS injection. Data are mean ± S.D. * P

    Journal: Journal of Cellular and Molecular Medicine

    Article Title: Progranulin deficiency leads to severe inflammation, lung injury and cell death in a mouse model of endotoxic shock

    doi: 10.1111/jcmm.12756

    Figure Lengend Snippet: PGRN deficiency increased the production of systemic and local pro‐inflammatory mediators after injection of LPS . ELISA of serum levels of inflammatory mediators including tumour necrosis factor α ( TNF ‐α) ( A ), interleukin 6 ( IL ‐6) ( B ), IL ‐10 ( C ) and monocyte chemoattractant protein 1 ( MCP ‐1) ( D ). ( E ) Western blot analysis of cold‐inducible RNA ‐binding protein ( CIRP ) and high mobility group box protein 1 ( HMGB 1) in the serum from WT and Grn −/− mice with LPS injection at 6 and 24 hrs. PS red, Ponceau S red staining. Real‐time RT ‐ PCR analysis of mRNA levels of TNF ‐α ( F ), IL ‐6 ( G ), IL ‐1β ( H ) and IL ‐10 ( I ) in the lung after LPS injection. Data are mean ± S.D. * P

    Article Snippet: Then, 1 μg of DNA‐free total RNA was reverse transcribed by use of a one‐step RT‐PCR kit (TaKaRa Bio, Shiga, Japan).

    Techniques: Injection, Enzyme-linked Immunosorbent Assay, Western Blot, RNA Binding Assay, Mouse Assay, Staining, Quantitative RT-PCR

    Overexpression of dominant negative NRF-1 attenuates the increase in β1 subunit mRNA levels in response to neuronal stimulation of primary cortical neurons. Primary cortical neurons were transfected with empty vector (pcDNA3) or dominant negative NRF-1 (NRF-1 DN; using Nucleofection™) and plated in 6-well plates. DIV7 cells were treated with either vehicle or 20 mM KCl for 6 h. Total mRNA was isolated from cells and quantified by TaqMan qRT-PCR. Gabrb1 mRNA expression was normalized to Cyclophilin A mRNA levels and is presented relative to its levels in pcDNA3 transfected neurons that were treated with vehicle (expressed as 1). Data represent the average ± SEM of n = 3 independent primary neuronal cultures. Two-way ANOVA was performed (comparison between treatments, p = 0.0148; comparison between transfected DNA conditions, p = 0.0031) with Bonferroni post hoc analysis (adjusted p value = 0.0263 for KCl vs. water with pcDNA transfection, not significant for NRF-1 DN).

    Journal: Frontiers in Molecular Neuroscience

    Article Title: Nuclear Respiratory Factor 1 (NRF-1) Controls the Activity Dependent Transcription of the GABA-A Receptor Beta 1 Subunit Gene in Neurons

    doi: 10.3389/fnmol.2018.00285

    Figure Lengend Snippet: Overexpression of dominant negative NRF-1 attenuates the increase in β1 subunit mRNA levels in response to neuronal stimulation of primary cortical neurons. Primary cortical neurons were transfected with empty vector (pcDNA3) or dominant negative NRF-1 (NRF-1 DN; using Nucleofection™) and plated in 6-well plates. DIV7 cells were treated with either vehicle or 20 mM KCl for 6 h. Total mRNA was isolated from cells and quantified by TaqMan qRT-PCR. Gabrb1 mRNA expression was normalized to Cyclophilin A mRNA levels and is presented relative to its levels in pcDNA3 transfected neurons that were treated with vehicle (expressed as 1). Data represent the average ± SEM of n = 3 independent primary neuronal cultures. Two-way ANOVA was performed (comparison between treatments, p = 0.0148; comparison between transfected DNA conditions, p = 0.0031) with Bonferroni post hoc analysis (adjusted p value = 0.0263 for KCl vs. water with pcDNA transfection, not significant for NRF-1 DN).

    Article Snippet: For each reaction, 20 ng of total RNA was reverse-transcribed to cDNA and PCR amplified in a single reaction mixture using the TaqMan® One-Step RT-PCR Master Mix Reagents Kit (Applied Biosystems).

    Techniques: Over Expression, Dominant Negative Mutation, Transfection, Plasmid Preparation, Isolation, Quantitative RT-PCR, Expressing

    NGF or BDNF changes the gene expression profile of BMSC and increases IL-6 mRNA. BMSC were treated with 100 ng/ml of (A) NGF or (B) BDNF for 18 hours. The cells were collected, RNA was isolated, and microarray analyses were performed. Graphs summarize the panel of gene expression changes in the BMSC treated with NGF or BDNF compared to untreated control cells. BMSC were treated with 100 ng/ml of (C) NGF or (D) BDNF for 2, 4 and 8 hours. The cells were collected, RNA was isolated and real time PCR was performed with the one-step QuantiTech SYBR Green kit as instructed by the manufacturer. BMSC IL-6 expression in response to NGF or BDNF exposure compared to untreated control cells is shown. Expression was normalized to the housekeeping gene GUSB . Statistical analysis was completed by ANOVA ( P ≤0.0001) with significance indicated by an asterisk.

    Journal: PLoS ONE

    Article Title: Neurotrophins Regulate Bone Marrow Stromal Cell IL-6 Expression through the MAPK Pathway

    doi: 10.1371/journal.pone.0009690

    Figure Lengend Snippet: NGF or BDNF changes the gene expression profile of BMSC and increases IL-6 mRNA. BMSC were treated with 100 ng/ml of (A) NGF or (B) BDNF for 18 hours. The cells were collected, RNA was isolated, and microarray analyses were performed. Graphs summarize the panel of gene expression changes in the BMSC treated with NGF or BDNF compared to untreated control cells. BMSC were treated with 100 ng/ml of (C) NGF or (D) BDNF for 2, 4 and 8 hours. The cells were collected, RNA was isolated and real time PCR was performed with the one-step QuantiTech SYBR Green kit as instructed by the manufacturer. BMSC IL-6 expression in response to NGF or BDNF exposure compared to untreated control cells is shown. Expression was normalized to the housekeeping gene GUSB . Statistical analysis was completed by ANOVA ( P ≤0.0001) with significance indicated by an asterisk.

    Article Snippet: The one-step QuantiTect SYBR Green RT-PCR Kit (Qiagen) was used as recommended by the manufacturer.

    Techniques: Expressing, Isolation, Microarray, Real-time Polymerase Chain Reaction, SYBR Green Assay

    Timing of non‐biologic product–associated cases of Theiler's disease and predicted exposure periods. Ten non‐biologic‐associated cases of Theiler's disease were diagnosed on 6 properties (properties A‐F) between January 2014 and February 2018. Each property is indicated by a different color. On 4 properties, there was follow‐up monitoring of in‐contact horses by examination, serum chemistry, and equine parvovirus‐hepatitis quantitative‐PCR. The time span during which new cases of hepatitis were identified on each farm is shown above the timeline (diamonds indicate that only 1 case occurred on that farm). In equine biologic product–associated cases, hepatitis typically occurs 4‐10 weeks after administration of an equine biologic product. Assuming a similar time frame between exposure and disease for non‐biologic‐associated cases, a predicted timeline of exposure for each farm was constructed, calculated as 10 weeks before the 1st case to 4 weeks before the last case

    Journal: Journal of Veterinary Internal Medicine

    Article Title: Viral testing of 10 cases of Theiler's disease and 37 in‐contact horses in the absence of equine biologic product administration: A prospective study (2014‐2018), et al. Viral testing of 10 cases of Theiler's disease and 37 in‐contact horses in the absence of equine biologic product administration: A prospective study (2014‐2018)

    doi: 10.1111/jvim.15362

    Figure Lengend Snippet: Timing of non‐biologic product–associated cases of Theiler's disease and predicted exposure periods. Ten non‐biologic‐associated cases of Theiler's disease were diagnosed on 6 properties (properties A‐F) between January 2014 and February 2018. Each property is indicated by a different color. On 4 properties, there was follow‐up monitoring of in‐contact horses by examination, serum chemistry, and equine parvovirus‐hepatitis quantitative‐PCR. The time span during which new cases of hepatitis were identified on each farm is shown above the timeline (diamonds indicate that only 1 case occurred on that farm). In equine biologic product–associated cases, hepatitis typically occurs 4‐10 weeks after administration of an equine biologic product. Assuming a similar time frame between exposure and disease for non‐biologic‐associated cases, a predicted timeline of exposure for each farm was constructed, calculated as 10 weeks before the 1st case to 4 weeks before the last case

    Article Snippet: All PCR mixtures used the Path ID multiplex RT‐qPCR kit (catalog no. 4442137; Thermo Fisher Scientific, Waltham, MA, USA) and 4 μL of extracted nucleic acids in a 25‐μL reaction volume.

    Techniques: Real-time Polymerase Chain Reaction, Construct

    HIV-2 Vpx represses TLR-mediated IL6, IL12p40, and TNFα induction. THP-1 cells infected for 48 h with equal amounts of HIV-2 viruses with or without vpx and pseudotyped with the VSVg envelope ( A and B ) were treated with TLR ligands for various times as indicated, and then total RNA was isolated, and mRNA levels were measured by qRT-PCR of IL6, IL12p40, and TNFα ( A ) or FLIP or IκBα, and β-actin (as an internal control) ( B ). Each mRNA induction was calculated based on untreated THP-1 cells. Each RT-PCR was run in triplicate. Data are shown as one representative example of three independent experiments, and values are mean ± S.D. C, total DNA was isolated from HIV-2-infected cells, and the level of viral DNA was determined by measuring Gag gene using real time qPCR.

    Journal: The Journal of Biological Chemistry

    Article Title: HIV-2 Vpx Protein Interacts with Interferon Regulatory Factor 5 (IRF5) and Inhibits Its Function *

    doi: 10.1074/jbc.M113.534321

    Figure Lengend Snippet: HIV-2 Vpx represses TLR-mediated IL6, IL12p40, and TNFα induction. THP-1 cells infected for 48 h with equal amounts of HIV-2 viruses with or without vpx and pseudotyped with the VSVg envelope ( A and B ) were treated with TLR ligands for various times as indicated, and then total RNA was isolated, and mRNA levels were measured by qRT-PCR of IL6, IL12p40, and TNFα ( A ) or FLIP or IκBα, and β-actin (as an internal control) ( B ). Each mRNA induction was calculated based on untreated THP-1 cells. Each RT-PCR was run in triplicate. Data are shown as one representative example of three independent experiments, and values are mean ± S.D. C, total DNA was isolated from HIV-2-infected cells, and the level of viral DNA was determined by measuring Gag gene using real time qPCR.

    Article Snippet: Real time qRT-PCR was performed using the one-step RT-PCR kit (Bio-Rad) to measure the mRNA level according to the manufacturer's instructions.

    Techniques: Infection, Isolation, Quantitative RT-PCR, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction

    HIV-2 Vpx represses TLR-mediated IL6, IL12p40, and TNFα induction. THP-1 cells transduced with an empty lentiviral vector or the vector encoding HIV-2 vpx ( A and B ) were treated with TLR ligands for various times as indicated, and then total RNA was isolated, and mRNA levels were measured by qRT-PCR of IL6, IL12p40, and TNFα ( A ) or FLIP or IκBα and β-actin (as an internal control) ( B ). Each mRNA induction was calculated based on untreated THP-1 cells. Each RT-PCR was run in triplicate. Data are shown as one representative example of three independent experiments, and values are mean ± S.D.

    Journal: The Journal of Biological Chemistry

    Article Title: HIV-2 Vpx Protein Interacts with Interferon Regulatory Factor 5 (IRF5) and Inhibits Its Function *

    doi: 10.1074/jbc.M113.534321

    Figure Lengend Snippet: HIV-2 Vpx represses TLR-mediated IL6, IL12p40, and TNFα induction. THP-1 cells transduced with an empty lentiviral vector or the vector encoding HIV-2 vpx ( A and B ) were treated with TLR ligands for various times as indicated, and then total RNA was isolated, and mRNA levels were measured by qRT-PCR of IL6, IL12p40, and TNFα ( A ) or FLIP or IκBα and β-actin (as an internal control) ( B ). Each mRNA induction was calculated based on untreated THP-1 cells. Each RT-PCR was run in triplicate. Data are shown as one representative example of three independent experiments, and values are mean ± S.D.

    Article Snippet: Real time qRT-PCR was performed using the one-step RT-PCR kit (Bio-Rad) to measure the mRNA level according to the manufacturer's instructions.

    Techniques: Transduction, Plasmid Preparation, Isolation, Quantitative RT-PCR, Reverse Transcription Polymerase Chain Reaction

    Relative expression of PKD isoforms in HeLa cells and their depletion by siRNA. (A) Analysis of mRNA expression by RT-PCR shows that PKD2 and PKD3 are the only PKD isoforms expressed in HeLa cells. RT-PCR reaction without the reverse transcriptase (RT−) was used as a negative control and PCR with the corresponding PKD cDNA (C) as a positive control. (B) Quantitative real-time RT-PCR analysis was performed on RNA extracted from HeLa cells. Bars represent the mean (±SD) of the relative mRNA expression of each PKD isoform compared with the average expression of β-actin. *, P

    Journal: The Journal of Cell Biology

    Article Title: Dimeric PKD regulates membrane fission to form transport carriers at the TGN

    doi: 10.1083/jcb.200703166

    Figure Lengend Snippet: Relative expression of PKD isoforms in HeLa cells and their depletion by siRNA. (A) Analysis of mRNA expression by RT-PCR shows that PKD2 and PKD3 are the only PKD isoforms expressed in HeLa cells. RT-PCR reaction without the reverse transcriptase (RT−) was used as a negative control and PCR with the corresponding PKD cDNA (C) as a positive control. (B) Quantitative real-time RT-PCR analysis was performed on RNA extracted from HeLa cells. Bars represent the mean (±SD) of the relative mRNA expression of each PKD isoform compared with the average expression of β-actin. *, P

    Article Snippet: RT-PCR Total RNA was extracted from HeLa cells with Nucleospin RNAII (Machery-Nagel) and reverse-transcribed with Titanium One-Step RT-PCR kit (Clontech Laboratories, Inc.) according to the manufacturer's instructions.

    Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Negative Control, Polymerase Chain Reaction, Positive Control, Quantitative RT-PCR

    Sensitivity analysis of three reverse primers used in the rRT-PCR assay. Cq values are shown against a ten-fold serial dilution of OROV RNA, either from the prototype strain, or an Ecuadorian strain.

    Journal: bioRxiv

    Article Title: Oropouche virus cases identified in Ecuador using an optimised rRT-PCR informed by metagenomic sequencing

    doi: 10.1101/683953

    Figure Lengend Snippet: Sensitivity analysis of three reverse primers used in the rRT-PCR assay. Cq values are shown against a ten-fold serial dilution of OROV RNA, either from the prototype strain, or an Ecuadorian strain.

    Article Snippet: rRT-PCR limit of detection analysisFor OROV rRT-PCR assay optimisation (supplementary material), a range of primers were tested (supplementary material, table S1) using Invitrogen Superscript III Platinum One-step Quantitative RT-PCR kit (Invitrogen) in a standard 25 µL reaction, run on an ABI 7500 real-time PCR machine (Applied Biosystems).

    Techniques: Quantitative RT-PCR, Serial Dilution