quantitative polymerase chain reaction qpcr  (Thermo Fisher)


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    Thermo Fisher quantitative polymerase chain reaction qpcr
    Ct values for M . <t>leprae</t> target (16S rRNA) amplified by <t>qPCR</t> from infected mouse footpad tissue extracted by different DNA extraction procedures. Data are displayed as mean plus standard deviation (SD) from batches processed independently. All kits were analyzed in triplicate with the exception of Microbiome, which was analyzed in duplicate.
    Quantitative Polymerase Chain Reaction Qpcr, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 340 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 92 stars, based on 340 article reviews
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    quantitative polymerase chain reaction qpcr - by Bioz Stars, 2020-09
    92/100 stars

    Images

    1) Product Images from "Ultra-sensitive detection of Mycobacterium leprae: DNA extraction and PCR assays"

    Article Title: Ultra-sensitive detection of Mycobacterium leprae: DNA extraction and PCR assays

    Journal: PLoS Neglected Tropical Diseases

    doi: 10.1371/journal.pntd.0008325

    Ct values for M . leprae target (16S rRNA) amplified by qPCR from infected mouse footpad tissue extracted by different DNA extraction procedures. Data are displayed as mean plus standard deviation (SD) from batches processed independently. All kits were analyzed in triplicate with the exception of Microbiome, which was analyzed in duplicate.
    Figure Legend Snippet: Ct values for M . leprae target (16S rRNA) amplified by qPCR from infected mouse footpad tissue extracted by different DNA extraction procedures. Data are displayed as mean plus standard deviation (SD) from batches processed independently. All kits were analyzed in triplicate with the exception of Microbiome, which was analyzed in duplicate.

    Techniques Used: Amplification, Real-time Polymerase Chain Reaction, Infection, DNA Extraction, Standard Deviation

    2) Product Images from "Loss of SYNJ1 dual phosphatase activity leads to early onset refractory seizures and progressive neurological decline"

    Article Title: Loss of SYNJ1 dual phosphatase activity leads to early onset refractory seizures and progressive neurological decline

    Journal: Brain

    doi: 10.1093/brain/aww180

    Levels of SYNJ1 mRNA in patient and control fibroblasts. The effect of the premature stop variants identified in Families B and C was evaluated by the amount of mutant mRNA transcript in patient fibroblasts. Messenger RNA was extracted from human fibroblasts expressing either homozygous wild-type (control) or mutant SYNJ1: patient Family B homozygous for p.Trp843* and patient Family C compound heterozygous for p.Gln647Argfs*6/p.Ser1122Thrfs*3. Quantitative PCR was performed on the converted cDNA. Graphics depict mean values of normalized ΔCt, relatively compared to wild-type mRNA levels, from at least two independent experiments (bars reflect standard error of mean). P -values were determined by comparing the normalized ΔCt (unpaired t -test, **** P
    Figure Legend Snippet: Levels of SYNJ1 mRNA in patient and control fibroblasts. The effect of the premature stop variants identified in Families B and C was evaluated by the amount of mutant mRNA transcript in patient fibroblasts. Messenger RNA was extracted from human fibroblasts expressing either homozygous wild-type (control) or mutant SYNJ1: patient Family B homozygous for p.Trp843* and patient Family C compound heterozygous for p.Gln647Argfs*6/p.Ser1122Thrfs*3. Quantitative PCR was performed on the converted cDNA. Graphics depict mean values of normalized ΔCt, relatively compared to wild-type mRNA levels, from at least two independent experiments (bars reflect standard error of mean). P -values were determined by comparing the normalized ΔCt (unpaired t -test, **** P

    Techniques Used: Mutagenesis, Expressing, Real-time Polymerase Chain Reaction

    3) Product Images from "Inhibition of microRNA-21 upregulates the expression of programmed cell death 4 and phosphatase tensin homologue in the A431 squamous cell carcinoma cell line"

    Article Title: Inhibition of microRNA-21 upregulates the expression of programmed cell death 4 and phosphatase tensin homologue in the A431 squamous cell carcinoma cell line

    Journal: Oncology Letters

    doi: 10.3892/ol.2014.2066

    (A) Quantitative polymerase chain reaction (qPCR) result demonstrating mRNA expression of miR-21 in A431 cells, cells transfected with an unrelated fragment of control (NC) and cells transfected with ASO-miR-21. (B) The relative expression level of miR-21 in the A431 cells was significantly decreased by ASO-miR-21 compared with the control (P
    Figure Legend Snippet: (A) Quantitative polymerase chain reaction (qPCR) result demonstrating mRNA expression of miR-21 in A431 cells, cells transfected with an unrelated fragment of control (NC) and cells transfected with ASO-miR-21. (B) The relative expression level of miR-21 in the A431 cells was significantly decreased by ASO-miR-21 compared with the control (P

    Techniques Used: Real-time Polymerase Chain Reaction, Expressing, Transfection, Allele-specific Oligonucleotide

    4) Product Images from "Exosomes from Plasmodium-infected hosts inhibit tumor angiogenesis in a murine Lewis lung cancer model"

    Article Title: Exosomes from Plasmodium-infected hosts inhibit tumor angiogenesis in a murine Lewis lung cancer model

    Journal: Oncogenesis

    doi: 10.1038/oncsis.2017.52

    miRNAs inhibitors rescued the effect of exosomes inhibition on VEGFR2 expression and tube formation in endothelial cells. ( a ) The relative expression of VEGFR2 mRNA in MS1 cells co-cultured with different groups exosomes and transfected with miRNAs inhibitors based on qPCR (*** P
    Figure Legend Snippet: miRNAs inhibitors rescued the effect of exosomes inhibition on VEGFR2 expression and tube formation in endothelial cells. ( a ) The relative expression of VEGFR2 mRNA in MS1 cells co-cultured with different groups exosomes and transfected with miRNAs inhibitors based on qPCR (*** P

    Techniques Used: Inhibition, Expressing, Cell Culture, Transfection, Real-time Polymerase Chain Reaction

    Exosomes inhibited vegfR2 expression in MS1 cells. ( a ) MS1 cells were cultured in vitro and exosomes were added in the culture medium for 24 h. VEGFR2 mRNA expression was detected using qPCR (* P
    Figure Legend Snippet: Exosomes inhibited vegfR2 expression in MS1 cells. ( a ) MS1 cells were cultured in vitro and exosomes were added in the culture medium for 24 h. VEGFR2 mRNA expression was detected using qPCR (* P

    Techniques Used: Expressing, Cell Culture, In Vitro, Real-time Polymerase Chain Reaction

    5) Product Images from "Comparison of Periodontal Ligament Cell Lines with Adenovirus- and Lentivirus-Mediated Human Telomerase Reverse Transcription Expression"

    Article Title: Comparison of Periodontal Ligament Cell Lines with Adenovirus- and Lentivirus-Mediated Human Telomerase Reverse Transcription Expression

    Journal: Human Gene Therapy Methods

    doi: 10.1089/hgtb.2018.184

    Relative hTERT expression levels in PDL cells after transduction. Human PDL cells were transduced with the indicated vector, and the relative expression levels were determined by quantitative polymerase chain reaction (qPCR). The values represent the mean of 4 trials, and the error bars indicate the standard deviation. *** p
    Figure Legend Snippet: Relative hTERT expression levels in PDL cells after transduction. Human PDL cells were transduced with the indicated vector, and the relative expression levels were determined by quantitative polymerase chain reaction (qPCR). The values represent the mean of 4 trials, and the error bars indicate the standard deviation. *** p

    Techniques Used: Expressing, Transduction, Plasmid Preparation, Real-time Polymerase Chain Reaction, Standard Deviation

    6) Product Images from "Doxorubicin combined with celecoxib inhibits tumor growth of medullary thyroid carcinoma in xenografted mice"

    Article Title: Doxorubicin combined with celecoxib inhibits tumor growth of medullary thyroid carcinoma in xenografted mice

    Journal: Oncology Letters

    doi: 10.3892/ol.2014.2050

    Expression of COX-2 and MDR1 in the tumor tissue was determined following treatment with CXB, DOX or a combination for 21 days. Expression of (A) COX-2 and (B) MDR1 mRNA in tumor tissue, as determined by qPCR. (C) Protein expression level of COX-2 and MDR1 in tumor tissue, as determined by western blot analysis. * P
    Figure Legend Snippet: Expression of COX-2 and MDR1 in the tumor tissue was determined following treatment with CXB, DOX or a combination for 21 days. Expression of (A) COX-2 and (B) MDR1 mRNA in tumor tissue, as determined by qPCR. (C) Protein expression level of COX-2 and MDR1 in tumor tissue, as determined by western blot analysis. * P

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction, Western Blot

    7) Product Images from "Flaxseed lignans enriched in secoisolariciresinol diglucoside prevent acute asbestos-induced peritoneal inflammation in mice"

    Article Title: Flaxseed lignans enriched in secoisolariciresinol diglucoside prevent acute asbestos-induced peritoneal inflammation in mice

    Journal: Carcinogenesis

    doi: 10.1093/carcin/bgv174

    Flaxseed lignan diet blunts proinflammatory and profibrogenic cytokine levels induced by asbestos exposure and alters cytokine and cytokine receptor gene expression. PLF levels (pg/ml) of proinflammatory TNFα ( A ) and profibrogenic free, active TGFß1 ( C ) were evaluated 3 days post asbestos exposure from mouse cohorts fed CTL or FLC diets. PLF WBC mRNA expression of cytokines, TNFα ( B ) and TGFß1 ( D ) and cytokine receptors, TNFαR1 ( E ) and TGFßR1 ( F ) was determined using qPCR. Levels of target gene mRNA were normalized to 18S ribosomal RNA and values are expressed as fold change from CTL. # indicates a statistically significant difference (#### P
    Figure Legend Snippet: Flaxseed lignan diet blunts proinflammatory and profibrogenic cytokine levels induced by asbestos exposure and alters cytokine and cytokine receptor gene expression. PLF levels (pg/ml) of proinflammatory TNFα ( A ) and profibrogenic free, active TGFß1 ( C ) were evaluated 3 days post asbestos exposure from mouse cohorts fed CTL or FLC diets. PLF WBC mRNA expression of cytokines, TNFα ( B ) and TGFß1 ( D ) and cytokine receptors, TNFαR1 ( E ) and TGFßR1 ( F ) was determined using qPCR. Levels of target gene mRNA were normalized to 18S ribosomal RNA and values are expressed as fold change from CTL. # indicates a statistically significant difference (#### P

    Techniques Used: Expressing, CTL Assay, Real-time Polymerase Chain Reaction

    Flaxseed lignan diet abrogates increased nitrosative and oxidative stress from exposure to asbestos fibers. Mouse cohorts (13 weeks old) were fed CTL ( n = 16) or FLC ( n = 17) diets for 7 days prior to exposure to 400 µg of crocidolite asbestos fibers. PLF was evaluated 3 days following asbestos exposure for nitrite ( A ) and MDA concentrations ( C ). PLF WBC mRNA expression of iNOS ( B ), HO-1 ( D ), Nqo1 ( E ) and Gstm1 ( F ) was determined using qPCR. Levels of target gene mRNA were normalized to 18S ribosomal RNA and values are expressed as fold change from CTL. # indicates a statistically significant difference (#### P
    Figure Legend Snippet: Flaxseed lignan diet abrogates increased nitrosative and oxidative stress from exposure to asbestos fibers. Mouse cohorts (13 weeks old) were fed CTL ( n = 16) or FLC ( n = 17) diets for 7 days prior to exposure to 400 µg of crocidolite asbestos fibers. PLF was evaluated 3 days following asbestos exposure for nitrite ( A ) and MDA concentrations ( C ). PLF WBC mRNA expression of iNOS ( B ), HO-1 ( D ), Nqo1 ( E ) and Gstm1 ( F ) was determined using qPCR. Levels of target gene mRNA were normalized to 18S ribosomal RNA and values are expressed as fold change from CTL. # indicates a statistically significant difference (#### P

    Techniques Used: CTL Assay, Multiple Displacement Amplification, Expressing, Real-time Polymerase Chain Reaction

    Flaxseed lignan diet ameliorates asbestos-induced WBC proinflammatory cytokine release and gene expression levels. PLF levels (pg/ml for IL-1ß and IL-6 and ng/ml for HMGB1) of proinflammatory cytokines IL-1ß ( A ), IL-6 ( C ) and HMGB1 ( E ) were evaluated 3 days post asbestos exposure. PLF WBC mRNA expression of IL-1ß ( B ), IL-6 ( D ) and HMGB1 ( F ) was determined using qPCR. Levels of target gene mRNA were normalized to 18S ribosomal RNA and values are expressed as fold change from CTL. # indicates a statistically significant difference (#### P
    Figure Legend Snippet: Flaxseed lignan diet ameliorates asbestos-induced WBC proinflammatory cytokine release and gene expression levels. PLF levels (pg/ml for IL-1ß and IL-6 and ng/ml for HMGB1) of proinflammatory cytokines IL-1ß ( A ), IL-6 ( C ) and HMGB1 ( E ) were evaluated 3 days post asbestos exposure. PLF WBC mRNA expression of IL-1ß ( B ), IL-6 ( D ) and HMGB1 ( F ) was determined using qPCR. Levels of target gene mRNA were normalized to 18S ribosomal RNA and values are expressed as fold change from CTL. # indicates a statistically significant difference (#### P

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction, CTL Assay

    8) Product Images from "BReast CAncer susceptibility gene 2 deficiency exacerbates oxidized LDL‐induced DNA damage and endothelial apoptosis, et al. BReast CAncer susceptibility gene 2 deficiency exacerbates oxidized LDL‐induced DNA damage and endothelial apoptosis"

    Article Title: BReast CAncer susceptibility gene 2 deficiency exacerbates oxidized LDL‐induced DNA damage and endothelial apoptosis, et al. BReast CAncer susceptibility gene 2 deficiency exacerbates oxidized LDL‐induced DNA damage and endothelial apoptosis

    Journal: Physiological Reports

    doi: 10.14814/phy2.14481

    Oxidized LDL induces increased expression of DNA damage‐related molecules, p53, and related apoptotic molecules in BRCA2‐deficient endothelial cells. Proteins were extracted from HUVECs transfected either with scrambled control or siBRCA2 for 48 hr and then treated with oxLDL (100 μg/ml) for 24 hr to perform immunoblot for (a) p(phospho)ATM, ATM, pCHK1, and CHK1, (b) p53, Bax, and Bcl‐2, (e) p21. GAPDH was used as a loading control. Protein quantification for p53 (c) and for the Bax/Bcl‐2 ratio (d). (f) qPCR for p21 . (g) Proliferation was evaluated in HUVECs transfected either with scrambled control or siBRCA2 for 48 hr and then treated with oxLDL for additional 24 hr. N = 3–4/group for immunoblot and qPCR in triplicates. Six‐wells/group for proliferation. *, ** and *** p
    Figure Legend Snippet: Oxidized LDL induces increased expression of DNA damage‐related molecules, p53, and related apoptotic molecules in BRCA2‐deficient endothelial cells. Proteins were extracted from HUVECs transfected either with scrambled control or siBRCA2 for 48 hr and then treated with oxLDL (100 μg/ml) for 24 hr to perform immunoblot for (a) p(phospho)ATM, ATM, pCHK1, and CHK1, (b) p53, Bax, and Bcl‐2, (e) p21. GAPDH was used as a loading control. Protein quantification for p53 (c) and for the Bax/Bcl‐2 ratio (d). (f) qPCR for p21 . (g) Proliferation was evaluated in HUVECs transfected either with scrambled control or siBRCA2 for 48 hr and then treated with oxLDL for additional 24 hr. N = 3–4/group for immunoblot and qPCR in triplicates. Six‐wells/group for proliferation. *, ** and *** p

    Techniques Used: Expressing, Transfection, Real-time Polymerase Chain Reaction

    OxLDL‐induced ROS, DNA damage, and apoptosis is exacerbated in BRCA2‐deficient endothelial cells. HUVECs were transfected either with scrambled control or siBRCA2, and RNA and protein were extracted 24, 48, and 72 hr post‐transfection to perform (a) qPCR and (b) immunoblotting for BRCA2. (c) ROS was measured in HUVECs after transfecting with either scrambled control or siBRCA2 for 48 hr and then treated with oxLDL (100 μg/ml) for 12 hr. Twelve‐wells/group. (d) Immunoblotting for γH2AX, H2AX, and cleaved caspase‐3 in HUVECs transfected with either scrambled control or siBRCA2 for 48 hr and then treated with oxLDL (100 μg/ml) for 24 hr. (e) quantification for the protein levels of cleaved caspase‐3. GAPDH was used as a loading control. (f) Apoptosis was also measured by flow cytometry in HUVECs transfected with either scrambled control or siBRCA2 for 48 hr and then treated with oxLDL (100 μg/ml) for 24 hr. N = 3–4 in triplicates. *, ** and *** p
    Figure Legend Snippet: OxLDL‐induced ROS, DNA damage, and apoptosis is exacerbated in BRCA2‐deficient endothelial cells. HUVECs were transfected either with scrambled control or siBRCA2, and RNA and protein were extracted 24, 48, and 72 hr post‐transfection to perform (a) qPCR and (b) immunoblotting for BRCA2. (c) ROS was measured in HUVECs after transfecting with either scrambled control or siBRCA2 for 48 hr and then treated with oxLDL (100 μg/ml) for 12 hr. Twelve‐wells/group. (d) Immunoblotting for γH2AX, H2AX, and cleaved caspase‐3 in HUVECs transfected with either scrambled control or siBRCA2 for 48 hr and then treated with oxLDL (100 μg/ml) for 24 hr. (e) quantification for the protein levels of cleaved caspase‐3. GAPDH was used as a loading control. (f) Apoptosis was also measured by flow cytometry in HUVECs transfected with either scrambled control or siBRCA2 for 48 hr and then treated with oxLDL (100 μg/ml) for 24 hr. N = 3–4 in triplicates. *, ** and *** p

    Techniques Used: Transfection, Real-time Polymerase Chain Reaction, Flow Cytometry

    9) Product Images from "Asbestos Induces Oxidative Stress and Activation of Nrf2 Signaling in Murine Macrophages: Chemopreventive Role of the Synthetic Lignan Secoisolariciresinol Diglucoside (LGM2605)"

    Article Title: Asbestos Induces Oxidative Stress and Activation of Nrf2 Signaling in Murine Macrophages: Chemopreventive Role of the Synthetic Lignan Secoisolariciresinol Diglucoside (LGM2605)

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms17030322

    LGM2605 Boosts Nrf2 Activation and the Expression of Antioxidant Enzymes in Elicited Murine Peritoneal Macrophages. ( a ) Levels of active, nuclear Nrf2 were determined at 0, 1, 2, 4, 6, 8, and 12-h post asbestos exposure; Macrophage mRNA expression of ( b ) HO-1 and ( c ) NQO1 was determined at 0, 8, and 24-h post asbestos exposure using qPCR. Levels of target gene mRNA were normalized to β-actin RNA and values are expressed as fold change from CTL. Data are presented as mean ± SEM. * indicates a statistically significant difference ( p
    Figure Legend Snippet: LGM2605 Boosts Nrf2 Activation and the Expression of Antioxidant Enzymes in Elicited Murine Peritoneal Macrophages. ( a ) Levels of active, nuclear Nrf2 were determined at 0, 1, 2, 4, 6, 8, and 12-h post asbestos exposure; Macrophage mRNA expression of ( b ) HO-1 and ( c ) NQO1 was determined at 0, 8, and 24-h post asbestos exposure using qPCR. Levels of target gene mRNA were normalized to β-actin RNA and values are expressed as fold change from CTL. Data are presented as mean ± SEM. * indicates a statistically significant difference ( p

    Techniques Used: Activation Assay, Expressing, Real-time Polymerase Chain Reaction, CTL Assay

    10) Product Images from "YAP and TAZ promote periosteal osteoblast precursor expansion and differentiation for fracture repair"

    Article Title: YAP and TAZ promote periosteal osteoblast precursor expansion and differentiation for fracture repair

    Journal: bioRxiv

    doi: 10.1101/2020.03.17.995761

    Adult onset-inducible, homozygous YAP/TAZ deletion from Osterix-expressing cells reduced callus bone formation. A) Micrographs of Picrosirius Red-stained calluses at 14 dpf. White boxes outline three regions of interest (B) in which second harmonic generated imaging (SHG) and anti-Osterix (αOSX) immunostaining are highlighted. Quantification of callus histomorphometry at 14 dpf of (C) the number Osterix-positive hypertrophic chondrocytes per cartilage area (OSX+ N.HC/CA), (D) Osterix-positive osteoblasts per bone surface (OSX+ N.Ob/BS), (E) bone area, and (F) percent bone area. Quantification of relative SHG intensity per bone area at 14 dpf of (G) the intact cortical bone, (H) the newly formed callus bone, and (I) normalized callus-to-intact SHG intensity. Messenger RNA was extracted from callus lysate preparations at 14 dpf and target gene expression was normalized to 18S rRNA and quantified as fold-change relative to wild type ( J-L ). J) Col10 and Vegfa mRNA expression. K) Col1a1, Col1a2 , and SerpinH1 mRNA expression. L) Runx2, Osx, Bsp , and Alp mRNA expression. Data are presented as individual samples in scatterplots and bars corresponding to the mean and standard error of the mean (SEM). N = 6 per group for qPCR and N = 3-4 per group for histomorphometry. Scale bars indicate 50 μm for all high-power images and 500 μm for callus images.
    Figure Legend Snippet: Adult onset-inducible, homozygous YAP/TAZ deletion from Osterix-expressing cells reduced callus bone formation. A) Micrographs of Picrosirius Red-stained calluses at 14 dpf. White boxes outline three regions of interest (B) in which second harmonic generated imaging (SHG) and anti-Osterix (αOSX) immunostaining are highlighted. Quantification of callus histomorphometry at 14 dpf of (C) the number Osterix-positive hypertrophic chondrocytes per cartilage area (OSX+ N.HC/CA), (D) Osterix-positive osteoblasts per bone surface (OSX+ N.Ob/BS), (E) bone area, and (F) percent bone area. Quantification of relative SHG intensity per bone area at 14 dpf of (G) the intact cortical bone, (H) the newly formed callus bone, and (I) normalized callus-to-intact SHG intensity. Messenger RNA was extracted from callus lysate preparations at 14 dpf and target gene expression was normalized to 18S rRNA and quantified as fold-change relative to wild type ( J-L ). J) Col10 and Vegfa mRNA expression. K) Col1a1, Col1a2 , and SerpinH1 mRNA expression. L) Runx2, Osx, Bsp , and Alp mRNA expression. Data are presented as individual samples in scatterplots and bars corresponding to the mean and standard error of the mean (SEM). N = 6 per group for qPCR and N = 3-4 per group for histomorphometry. Scale bars indicate 50 μm for all high-power images and 500 μm for callus images.

    Techniques Used: Expressing, Staining, Generated, Imaging, Immunostaining, Real-time Polymerase Chain Reaction

    11) Product Images from "Homemade-device-induced negative pressure promotes wound healing more efficiently than VSD-induced positive pressure by regulating inflammation, proliferation and remodeling"

    Article Title: Homemade-device-induced negative pressure promotes wound healing more efficiently than VSD-induced positive pressure by regulating inflammation, proliferation and remodeling

    Journal: International Journal of Molecular Medicine

    doi: 10.3892/ijmm.2017.2919

    Analysis of inflammatory and growth factor secretion in wound tissue in the NP and PP groups. (A) Representative images of H E staining of inflammatory cells in wound tissues following pressure treatment on days 3, 7, 10, and 14. The upper lane indicates the control group, the middle lane indicates the NP group, and the lower lane indicates the PP group. (B) Statistical quantification of macrophage infiltration in wound tissue at different time points. (C) Statistical quantification of neutrophil infiltration in wound tissue at different time points. Statistical analysis of the mRNA expression of (D) IL-1β, (E) IL-10, (F) EGF, (G) TGF-β1, (H) IGF-1, (I) bFGF, (J) PDGF, (K) VEGF in wound tissue examined by qPCR. Data are presented as the means ± SEM, n=3 experiments. *** P
    Figure Legend Snippet: Analysis of inflammatory and growth factor secretion in wound tissue in the NP and PP groups. (A) Representative images of H E staining of inflammatory cells in wound tissues following pressure treatment on days 3, 7, 10, and 14. The upper lane indicates the control group, the middle lane indicates the NP group, and the lower lane indicates the PP group. (B) Statistical quantification of macrophage infiltration in wound tissue at different time points. (C) Statistical quantification of neutrophil infiltration in wound tissue at different time points. Statistical analysis of the mRNA expression of (D) IL-1β, (E) IL-10, (F) EGF, (G) TGF-β1, (H) IGF-1, (I) bFGF, (J) PDGF, (K) VEGF in wound tissue examined by qPCR. Data are presented as the means ± SEM, n=3 experiments. *** P

    Techniques Used: Staining, Expressing, Real-time Polymerase Chain Reaction

    12) Product Images from "MicroRNA-1 Participates in Nitric Oxide-Induced Apoptotic Insults to MC3T3-E1 Cells by Targeting Heat-Shock Protein-70"

    Article Title: MicroRNA-1 Participates in Nitric Oxide-Induced Apoptotic Insults to MC3T3-E1 Cells by Targeting Heat-Shock Protein-70

    Journal: International Journal of Biological Sciences

    doi: 10.7150/ijbs.11138

    Roles of microRNA-1 (miR-1) in sodium nitroprusside (SNP)-induced inhibition of heat shock protein (HSP)-70 mRNA and protein expressions. A bioinformatic approach was conducted to predict the existence of miR-1-specific binding elements in the 3'-UTR of HSP-70 mRNA (A). MC3T3-E1 cells were treated with SNP, hsa-miR-1 (an miR-1 inhibitor), and their combination for 12 h. The Anti-miR™ miRNA Inhibitor Negative Control #1 (Control) was transfected into MC3T3-E1 cells as a negative control. Amounts of HSP-70 were immunodetected (B, top panel). β-Actin was analyzed as the internal control (bottom panel). These immunorelated protein bands were quantified and statistically analyzed (C). Levels of HSP-70 mRNA were quantified using a quantitative PCR (D). Each value represents the mean ± SEM, n = 6. * and # Values significantly ( p
    Figure Legend Snippet: Roles of microRNA-1 (miR-1) in sodium nitroprusside (SNP)-induced inhibition of heat shock protein (HSP)-70 mRNA and protein expressions. A bioinformatic approach was conducted to predict the existence of miR-1-specific binding elements in the 3'-UTR of HSP-70 mRNA (A). MC3T3-E1 cells were treated with SNP, hsa-miR-1 (an miR-1 inhibitor), and their combination for 12 h. The Anti-miR™ miRNA Inhibitor Negative Control #1 (Control) was transfected into MC3T3-E1 cells as a negative control. Amounts of HSP-70 were immunodetected (B, top panel). β-Actin was analyzed as the internal control (bottom panel). These immunorelated protein bands were quantified and statistically analyzed (C). Levels of HSP-70 mRNA were quantified using a quantitative PCR (D). Each value represents the mean ± SEM, n = 6. * and # Values significantly ( p

    Techniques Used: Inhibition, Binding Assay, Negative Control, Transfection, Real-time Polymerase Chain Reaction

    Effects of sodium nitroprusside (SNP) on microRNA-1 (miR-1) expression. MC3T3- E1 cells were exposed to 2 mM SNP for 1, 3, 6, 12, and 24 h. Levels of miR-1 were quantified using a quantitative PCR analysis (A). MC3T3-E1 cells were treated with SNP, hsa-miR-1 (a miR-1 inhibitor), and their combination for 6 h. The Anti-miR™ miRNA Inhibitor Negative Control #1 (Control) was transfected into MC3T3-E1 cells as a negative control. Amounts of miR-1 were quantified using a quantitative PCR analysis (B). Each value represents the mean ± SEM, n = 6. * and # Values significantly ( p
    Figure Legend Snippet: Effects of sodium nitroprusside (SNP) on microRNA-1 (miR-1) expression. MC3T3- E1 cells were exposed to 2 mM SNP for 1, 3, 6, 12, and 24 h. Levels of miR-1 were quantified using a quantitative PCR analysis (A). MC3T3-E1 cells were treated with SNP, hsa-miR-1 (a miR-1 inhibitor), and their combination for 6 h. The Anti-miR™ miRNA Inhibitor Negative Control #1 (Control) was transfected into MC3T3-E1 cells as a negative control. Amounts of miR-1 were quantified using a quantitative PCR analysis (B). Each value represents the mean ± SEM, n = 6. * and # Values significantly ( p

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction, Negative Control, Transfection

    13) Product Images from "The effect of resveratrol on lifespan depends on both gender and dietary nutrient composition in Drosophila melanogaster"

    Article Title: The effect of resveratrol on lifespan depends on both gender and dietary nutrient composition in Drosophila melanogaster

    Journal: Age

    doi: 10.1007/s11357-011-9332-3

    The effect of resveratrol on gene expression in Canton S female flies fed the low sugar–high protein (LS–HP) diet as determined by quantitative PCR (qPCR). a Expression patterns of metabolism-related genes in flies fed the LS–HP
    Figure Legend Snippet: The effect of resveratrol on gene expression in Canton S female flies fed the low sugar–high protein (LS–HP) diet as determined by quantitative PCR (qPCR). a Expression patterns of metabolism-related genes in flies fed the LS–HP

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction

    14) Product Images from "Myeloid Neoplasia: Identification and targeting of novel CDK9 complexes in acute myeloid leukemia"

    Article Title: Myeloid Neoplasia: Identification and targeting of novel CDK9 complexes in acute myeloid leukemia

    Journal: Blood

    doi: 10.1182/blood-2018-08-870089

    The CDK9/mLST8/RAPTOR nuclear complex regulates expression of genes important for leukemogenesis. (A) MV4-11 cells were crosslinked with 1% formaldehyde. Chromatin-protein complexes were immunoprecipitated with anti-CDK9, anti-mLST8, and anti-RAPTOR antibodies. Rabbit IgG and anti-RICTOR antibodies were used as negative controls. qPCR was performed on immunoprecipitated DNA with primers for the MYC and PIM1 promoters. Data are expressed as fold enrichment over the IgG control. Shown are means + standard error (SE) of 4 independent experiments. (B) Cells were treated with atuveciclib for 0, 1, 2, and 4 hours. Cells were then lysed, and proteins were resolved by SDS-PAGE, followed by transfer to PVDF membranes. Membranes were immunoblotted with the indicated antibodies. The immunoblots with antibodies against the phosphorylated forms of the proteins or against the total proteins were from lysates from the same experiments analyzed in parallel by SDS-PAGE. (C) Cells were treated with control (DMSO) or atuveciclib for 4 hours and lysed, and proteins resolved by SDS-PAGE followed by transfer to polyvinylidene fluoride membranes. Membranes were then immunoblotted with the indicated antibodies. *Nonspecific band in the c-MYC blot. (D) qRT-PCR analysis of the relative mRNA expression of MYC and PIM1 in U937 cells after atuveciclib treatment of 4 hours. GAPDH was used for normalization. Data are expressed as fold change over the control. Shown are means + SE of 3 independent experiments. ** P
    Figure Legend Snippet: The CDK9/mLST8/RAPTOR nuclear complex regulates expression of genes important for leukemogenesis. (A) MV4-11 cells were crosslinked with 1% formaldehyde. Chromatin-protein complexes were immunoprecipitated with anti-CDK9, anti-mLST8, and anti-RAPTOR antibodies. Rabbit IgG and anti-RICTOR antibodies were used as negative controls. qPCR was performed on immunoprecipitated DNA with primers for the MYC and PIM1 promoters. Data are expressed as fold enrichment over the IgG control. Shown are means + standard error (SE) of 4 independent experiments. (B) Cells were treated with atuveciclib for 0, 1, 2, and 4 hours. Cells were then lysed, and proteins were resolved by SDS-PAGE, followed by transfer to PVDF membranes. Membranes were immunoblotted with the indicated antibodies. The immunoblots with antibodies against the phosphorylated forms of the proteins or against the total proteins were from lysates from the same experiments analyzed in parallel by SDS-PAGE. (C) Cells were treated with control (DMSO) or atuveciclib for 4 hours and lysed, and proteins resolved by SDS-PAGE followed by transfer to polyvinylidene fluoride membranes. Membranes were then immunoblotted with the indicated antibodies. *Nonspecific band in the c-MYC blot. (D) qRT-PCR analysis of the relative mRNA expression of MYC and PIM1 in U937 cells after atuveciclib treatment of 4 hours. GAPDH was used for normalization. Data are expressed as fold change over the control. Shown are means + SE of 3 independent experiments. ** P

    Techniques Used: Expressing, Immunoprecipitation, Real-time Polymerase Chain Reaction, SDS Page, Western Blot, Quantitative RT-PCR

    15) Product Images from "CXCR7 functions in colon cancer cell survival and migration"

    Article Title: CXCR7 functions in colon cancer cell survival and migration

    Journal: Experimental and Therapeutic Medicine

    doi: 10.3892/etm.2015.2748

    qPCR analysis of C-X-C chemokine receptor 7 mRNA expression in HT-29 and SW-480 cells. qPCR, quantitative polymerase chain reaction.
    Figure Legend Snippet: qPCR analysis of C-X-C chemokine receptor 7 mRNA expression in HT-29 and SW-480 cells. qPCR, quantitative polymerase chain reaction.

    Techniques Used: Real-time Polymerase Chain Reaction, Expressing

    16) Product Images from "A novel anti-mycobacterial function of mitogen-activated protein kinase phosphatase-1"

    Article Title: A novel anti-mycobacterial function of mitogen-activated protein kinase phosphatase-1

    Journal: BMC Immunology

    doi: 10.1186/1471-2172-10-64

    BCG induces MKP-1 expression . (A) Human monocytes were treated with BCG (MOI = 1 CFU/cell) for the indicated time points and RNA was harvested. MKP-1 level was assayed by using QPCR. Results are shown as mean ± SD from 3 different donors. * = p
    Figure Legend Snippet: BCG induces MKP-1 expression . (A) Human monocytes were treated with BCG (MOI = 1 CFU/cell) for the indicated time points and RNA was harvested. MKP-1 level was assayed by using QPCR. Results are shown as mean ± SD from 3 different donors. * = p

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction

    BCG-induced MKP-1 expression is dependent on p38 MAPK and ERK1/2 . Human monocytes were pretreated with several inhibitors including PD98059 (20 μM), SB203580 (100 nM), SP600125 (100 nM), and CAPE (5 μg/ml) for 1 hour. BCG (MOI = 1 CFU/cell) was then added for 1 hour and RNA was harvested. Levels of MKP-1 were measured by QPCR. Percentage change was defined as the percentage of the fold induction of (BCG + inhibitor) over the fold induction of BCG without the inhibitor. Sample of BCG was set as 100% for comparisons. Independent experiments were performed on blood cells from 4 different donors and the results are shown as mean ± SD. * = p
    Figure Legend Snippet: BCG-induced MKP-1 expression is dependent on p38 MAPK and ERK1/2 . Human monocytes were pretreated with several inhibitors including PD98059 (20 μM), SB203580 (100 nM), SP600125 (100 nM), and CAPE (5 μg/ml) for 1 hour. BCG (MOI = 1 CFU/cell) was then added for 1 hour and RNA was harvested. Levels of MKP-1 were measured by QPCR. Percentage change was defined as the percentage of the fold induction of (BCG + inhibitor) over the fold induction of BCG without the inhibitor. Sample of BCG was set as 100% for comparisons. Independent experiments were performed on blood cells from 4 different donors and the results are shown as mean ± SD. * = p

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction

    Transfection of MKP-1 siRNA into primary human blood monocytes can increase LPS-induced TNF-α level while decrease BCG-induced TNF-α expression . Cells were first transfected with control (ctrl) or MKP-1 siRNA (MKP-1 si, 200 nM) for 24 hours and then treated with different inducers as indicated. (A) After transfection, cells were treated with Mock (M) or BCG (B, MOI = 1 CFU/cell) for 3 hours. Levels of MKP-1 mRNA were measured by RT-PCR. (B) Monocytes were treated as in (A) for the indicated time points and MKP-1 proteins were measured by Western blotting. (C) Cells were treated with LPS (50 ng/ml) for the indicated time points. RNA was harvested, and levels of MKP-1 and TNF-α were measured by RT-PCR. For both (B) and (C), the intensities of the PCR/protein bands were determined by using Bio-Rad Quantity One imaging software. The intensities of bands were normalized to the corresponding control. The values in parenthesis are the relative normalized intensities compared to those of the control siRNA-transfected cells without other treatment. (D) BCG (MOI = 1 CFU/cell) was added for 3 hours. RNA was harvested and levels of TNF-α, IL-6 and IL-10 were measured by QPCR. Percentage change was defined as the percentage of the fold induction of (MKP-1 si + BCG) over the fold induction of (ctrl + BCG). Results of (ctrl + BCG) were set as 100%. For (A)-(C), independent experiments were done on monocytes from 3 different donors and one representative set of results is shown. For (D), experiments were performed on 4 different donors and the results are shown as mean ± SD. * = p
    Figure Legend Snippet: Transfection of MKP-1 siRNA into primary human blood monocytes can increase LPS-induced TNF-α level while decrease BCG-induced TNF-α expression . Cells were first transfected with control (ctrl) or MKP-1 siRNA (MKP-1 si, 200 nM) for 24 hours and then treated with different inducers as indicated. (A) After transfection, cells were treated with Mock (M) or BCG (B, MOI = 1 CFU/cell) for 3 hours. Levels of MKP-1 mRNA were measured by RT-PCR. (B) Monocytes were treated as in (A) for the indicated time points and MKP-1 proteins were measured by Western blotting. (C) Cells were treated with LPS (50 ng/ml) for the indicated time points. RNA was harvested, and levels of MKP-1 and TNF-α were measured by RT-PCR. For both (B) and (C), the intensities of the PCR/protein bands were determined by using Bio-Rad Quantity One imaging software. The intensities of bands were normalized to the corresponding control. The values in parenthesis are the relative normalized intensities compared to those of the control siRNA-transfected cells without other treatment. (D) BCG (MOI = 1 CFU/cell) was added for 3 hours. RNA was harvested and levels of TNF-α, IL-6 and IL-10 were measured by QPCR. Percentage change was defined as the percentage of the fold induction of (MKP-1 si + BCG) over the fold induction of (ctrl + BCG). Results of (ctrl + BCG) were set as 100%. For (A)-(C), independent experiments were done on monocytes from 3 different donors and one representative set of results is shown. For (D), experiments were performed on 4 different donors and the results are shown as mean ± SD. * = p

    Techniques Used: Transfection, Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Polymerase Chain Reaction, Imaging, Software, Real-time Polymerase Chain Reaction

    Different inducers show differential induction of MKP-1 . (A) Primary human blood monocytes were stimulated with LPS (50 ng/ml), Pam3Cys (50 ng/ml), or Poly IC (100 μg/ml) for 1 (open bars) and 3 (black bars) hours. Phosphate buffered saline (PBS) was used as a reagent control. RNA was harvested and MKP-1 mRNA was measured by QPCR method. (B) Primary human monocytes were treated with LPS (50 ng/ml) for the indicated time points and RNA samples were harvested. MKP-1 levels were studied by using QPCR. Independent experiments were done on blood cells from 4 different donors, and the results are shown as mean ± SD. * = p
    Figure Legend Snippet: Different inducers show differential induction of MKP-1 . (A) Primary human blood monocytes were stimulated with LPS (50 ng/ml), Pam3Cys (50 ng/ml), or Poly IC (100 μg/ml) for 1 (open bars) and 3 (black bars) hours. Phosphate buffered saline (PBS) was used as a reagent control. RNA was harvested and MKP-1 mRNA was measured by QPCR method. (B) Primary human monocytes were treated with LPS (50 ng/ml) for the indicated time points and RNA samples were harvested. MKP-1 levels were studied by using QPCR. Independent experiments were done on blood cells from 4 different donors, and the results are shown as mean ± SD. * = p

    Techniques Used: Real-time Polymerase Chain Reaction

    17) Product Images from "An exploratory study into the role of miR-204-5p in pregnancy-induced hypertension"

    Article Title: An exploratory study into the role of miR-204-5p in pregnancy-induced hypertension

    Journal: Experimental and Therapeutic Medicine

    doi: 10.3892/etm.2017.4212

    Expression of miRNAs in serum samples of healthy controls and patients with PIH, detected using reverse transcription-quantitative polymerase chain reaction. Expression of (A) miR-197-3p, (B) miR-92b-3p, (C) miR-342-3p, (D) miR-26a-5p, (E) miR-198, (F) miR-204-5p, (G) miR-296-5p and (H) miR-95-5p in serum samples of healthy normal controls and patients with PIH. Data are presented as the mean ± standard deviation. All experiments were repeated independently three times. ***P
    Figure Legend Snippet: Expression of miRNAs in serum samples of healthy controls and patients with PIH, detected using reverse transcription-quantitative polymerase chain reaction. Expression of (A) miR-197-3p, (B) miR-92b-3p, (C) miR-342-3p, (D) miR-26a-5p, (E) miR-198, (F) miR-204-5p, (G) miR-296-5p and (H) miR-95-5p in serum samples of healthy normal controls and patients with PIH. Data are presented as the mean ± standard deviation. All experiments were repeated independently three times. ***P

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction, Standard Deviation

    Related Articles

    Real-time Polymerase Chain Reaction:

    Article Title: Neurotrophin/Trk receptor signaling mediates C/EBP?, -? and NeuroD recruitment to immediate-early gene promoters in neuronal cells and requires C/EBPs to induce immediate-early gene transcription
    Article Snippet: .. Quantitative PCR qPCR or qRT-PCR was performed using the DyNAmoSYBR Green qPCR kit according to the manufacturer's protocol (Finnzymes, Milan, Italy) using the primers indicated in Table . .. For the ChIP, serial dilutions of IP DNA were subjected to real time PCR.

    Article Title: The Nuclear Protein Sge1 of Fusarium oxysporum Is Required for Parasitic Growth
    Article Snippet: .. Quantitative PCR (qPCR) was performed with Platinum SYBR Green qPCR (Invitrogen) using a 7500 Realtime PCR System (Applied Biosystems). ..

    Article Title: Phenotyping acute and chronic atopic dermatitis-like lesions in Stat6VT mice identifies a role for IL-33 in disease pathogenesis
    Article Snippet: .. Quantitative PCR (qPCR) was performed using TaqMan chemistry and a 7500 Fast Real-Time PCR System (Applied Biosystems). .. Primer and probe mixtures for Il4 (Mm00445259_m1), Il5 (Mm00439646_m1), Il13 (Mm00434204_m1), Il33 (Mm00505403_m1), Ifng (Mm01168134_m1), Tslp (Mm01157588_m1), B2m (Mm00437762_m1) and GAPDH ( Mm99999915_g1) were purchased from Applied Biosystems.

    Article Title: Elimination of “kitome” and “splashome” contamination results in lack of detection of a unique placental microbiome
    Article Snippet: .. 16S rRNA gene qPCRTo quantify bacterial loads in the communities, quantitative PCR (qPCR) was performed using 16S rRNA gene universal primers 357F (5′-CTCCTACGGGAGGCAGCAG-3′) and 519R (536R) (5′-GWATTACCGCGGCKGCTG-3′) [ ]. .. Reactions with SsoAdvanced™ Universal SYBR™Green Supermix (BioRad) were performed in triplicate in a 15 μl reaction, using 1:2 dilutions of DNA template.

    Article Title: Ultra-sensitive detection of Mycobacterium leprae: DNA extraction and PCR assays
    Article Snippet: .. Quantitative Polymerase Chain Reaction (qPCR) The levels of M . leprae from mouse footpads and clinical samples were estimated through the amplification of the M . leprae -specific 16S rRNA target [ ] in the TaqMan assay (Thermo Fisher Scientific). .. Another primer pair, which amplified a fragment of the mammalian 18S rRNA region, was used as an internal reference host DNA control (forward, 5´GAA ACT GCG AAT GGC TCA TTA AAT CA3´; reverse, 5´CCC GTC GGC ATG TAT TAG CTC T-3´; TaqMan probe, 5´TGG TTC CTT TGG TCG CTC GCT CC-3´).

    Article Title: Obesity promotes the expansion of metastasis-initiating cells in breast cancer
    Article Snippet: .. Complementary DNAs (cDNAs) were generated using oligo-T priming and the M-MLV transcriptase (H-) point mutant (Promega) and quantitative PCR (qPCR) was performed in a StepOnePlus thermocycler (Applied Biosystems) using the SYBR green PCR Master Mix (Kapa). ..

    Article Title: Viral DNA-Dependent Induction of Innate Immune Response to Hepatitis B Virus in Immortalized Mouse Hepatocytes
    Article Snippet: .. Quantitative PCR (qPCR) was performed on three technical replicates in adhesive-sealed 384-well plates on a 12K Flex machine (Life Technologies) in 10-μl volumes consisting of 1× Power SYBR green master mix or 1× TaqMan gene expression master mix (Life Technologies), 200 nM each primer for SYBR green or 150 nM each primer with 250 nM probe for TaqMan, and 100 ng cDNA. .. The data were validated by verifying specificity in a melting curve analysis for SYBR green experiments or by running several PCR products on a 3% agarose gel for TaqMan experiments.

    Article Title: Identification of limb-specific Lmx1b auto-regulatory modules with Nail-Patella Syndrome pathogenicity
    Article Snippet: .. Quantitative PCR (qPCR) was performed on the Mx3005P cycler using SYBRGreen PCR Master Mix (Life Technologies). .. Data were analyzed using the MxPro software (Agilent Technologies).

    Amplification:

    Article Title: Ultra-sensitive detection of Mycobacterium leprae: DNA extraction and PCR assays
    Article Snippet: .. Quantitative Polymerase Chain Reaction (qPCR) The levels of M . leprae from mouse footpads and clinical samples were estimated through the amplification of the M . leprae -specific 16S rRNA target [ ] in the TaqMan assay (Thermo Fisher Scientific). .. Another primer pair, which amplified a fragment of the mammalian 18S rRNA region, was used as an internal reference host DNA control (forward, 5´GAA ACT GCG AAT GGC TCA TTA AAT CA3´; reverse, 5´CCC GTC GGC ATG TAT TAG CTC T-3´; TaqMan probe, 5´TGG TTC CTT TGG TCG CTC GCT CC-3´).

    Mutagenesis:

    Article Title: Obesity promotes the expansion of metastasis-initiating cells in breast cancer
    Article Snippet: .. Complementary DNAs (cDNAs) were generated using oligo-T priming and the M-MLV transcriptase (H-) point mutant (Promega) and quantitative PCR (qPCR) was performed in a StepOnePlus thermocycler (Applied Biosystems) using the SYBR green PCR Master Mix (Kapa). ..

    Quantitative RT-PCR:

    Article Title: Neurotrophin/Trk receptor signaling mediates C/EBP?, -? and NeuroD recruitment to immediate-early gene promoters in neuronal cells and requires C/EBPs to induce immediate-early gene transcription
    Article Snippet: .. Quantitative PCR qPCR or qRT-PCR was performed using the DyNAmoSYBR Green qPCR kit according to the manufacturer's protocol (Finnzymes, Milan, Italy) using the primers indicated in Table . .. For the ChIP, serial dilutions of IP DNA were subjected to real time PCR.

    SYBR Green Assay:

    Article Title: The Nuclear Protein Sge1 of Fusarium oxysporum Is Required for Parasitic Growth
    Article Snippet: .. Quantitative PCR (qPCR) was performed with Platinum SYBR Green qPCR (Invitrogen) using a 7500 Realtime PCR System (Applied Biosystems). ..

    Article Title: Obesity promotes the expansion of metastasis-initiating cells in breast cancer
    Article Snippet: .. Complementary DNAs (cDNAs) were generated using oligo-T priming and the M-MLV transcriptase (H-) point mutant (Promega) and quantitative PCR (qPCR) was performed in a StepOnePlus thermocycler (Applied Biosystems) using the SYBR green PCR Master Mix (Kapa). ..

    Article Title: Viral DNA-Dependent Induction of Innate Immune Response to Hepatitis B Virus in Immortalized Mouse Hepatocytes
    Article Snippet: .. Quantitative PCR (qPCR) was performed on three technical replicates in adhesive-sealed 384-well plates on a 12K Flex machine (Life Technologies) in 10-μl volumes consisting of 1× Power SYBR green master mix or 1× TaqMan gene expression master mix (Life Technologies), 200 nM each primer for SYBR green or 150 nM each primer with 250 nM probe for TaqMan, and 100 ng cDNA. .. The data were validated by verifying specificity in a melting curve analysis for SYBR green experiments or by running several PCR products on a 3% agarose gel for TaqMan experiments.

    Generated:

    Article Title: Obesity promotes the expansion of metastasis-initiating cells in breast cancer
    Article Snippet: .. Complementary DNAs (cDNAs) were generated using oligo-T priming and the M-MLV transcriptase (H-) point mutant (Promega) and quantitative PCR (qPCR) was performed in a StepOnePlus thermocycler (Applied Biosystems) using the SYBR green PCR Master Mix (Kapa). ..

    Expressing:

    Article Title: Viral DNA-Dependent Induction of Innate Immune Response to Hepatitis B Virus in Immortalized Mouse Hepatocytes
    Article Snippet: .. Quantitative PCR (qPCR) was performed on three technical replicates in adhesive-sealed 384-well plates on a 12K Flex machine (Life Technologies) in 10-μl volumes consisting of 1× Power SYBR green master mix or 1× TaqMan gene expression master mix (Life Technologies), 200 nM each primer for SYBR green or 150 nM each primer with 250 nM probe for TaqMan, and 100 ng cDNA. .. The data were validated by verifying specificity in a melting curve analysis for SYBR green experiments or by running several PCR products on a 3% agarose gel for TaqMan experiments.

    Polymerase Chain Reaction:

    Article Title: The Nuclear Protein Sge1 of Fusarium oxysporum Is Required for Parasitic Growth
    Article Snippet: .. Quantitative PCR (qPCR) was performed with Platinum SYBR Green qPCR (Invitrogen) using a 7500 Realtime PCR System (Applied Biosystems). ..

    Article Title: Obesity promotes the expansion of metastasis-initiating cells in breast cancer
    Article Snippet: .. Complementary DNAs (cDNAs) were generated using oligo-T priming and the M-MLV transcriptase (H-) point mutant (Promega) and quantitative PCR (qPCR) was performed in a StepOnePlus thermocycler (Applied Biosystems) using the SYBR green PCR Master Mix (Kapa). ..

    Article Title: Identification of limb-specific Lmx1b auto-regulatory modules with Nail-Patella Syndrome pathogenicity
    Article Snippet: .. Quantitative PCR (qPCR) was performed on the Mx3005P cycler using SYBRGreen PCR Master Mix (Life Technologies). .. Data were analyzed using the MxPro software (Agilent Technologies).

    TaqMan Assay:

    Article Title: Ultra-sensitive detection of Mycobacterium leprae: DNA extraction and PCR assays
    Article Snippet: .. Quantitative Polymerase Chain Reaction (qPCR) The levels of M . leprae from mouse footpads and clinical samples were estimated through the amplification of the M . leprae -specific 16S rRNA target [ ] in the TaqMan assay (Thermo Fisher Scientific). .. Another primer pair, which amplified a fragment of the mammalian 18S rRNA region, was used as an internal reference host DNA control (forward, 5´GAA ACT GCG AAT GGC TCA TTA AAT CA3´; reverse, 5´CCC GTC GGC ATG TAT TAG CTC T-3´; TaqMan probe, 5´TGG TTC CTT TGG TCG CTC GCT CC-3´).

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    Thermo Fisher 18s qpcr target
    Droplet digital PCR for oocyst genome quantification. a One-dimensional scatter plots showing <t>18S</t> ddPCR assay on positive (syn18s standards from 7.2 × 10 4 to 0.72 copies/µL) and negative human blood (“HGD”) extracts. Clear demarcation between positive and negative partitions is shown. Uninfected human blood extract reaction was used to determine a universal positive/negative threshold set at 1853. b Quantification of syn18s standards using ddPCR in triplicate (black circles indicate the median of each run with error bars showing 95% CI) compared to the predicted <t>qPCR</t> value (grey line). c One-dimensional scatter plots showing 18S ddPCR assay on 14 oocyst-positive midguts, with positive and negative partitions. Uninfected mosquito midguts (“Neg”) were used to determine a universal positive/negative threshold set at 1853. d Quantification of genomes per oocyst for the14 microscopy-confirmed oocyst-positive midguts using 18S qPCR and 18S ddPCR. Box plots indicate the median and whiskers show the minimum and maximum responses. Groups compared using Wilcoxon matched-pairs signed rank test ( p = 0.43)
    18s Qpcr Target, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher real time quantitative polymerase chain reaction qpcr
    Effect of the anti‐human C5 antibody eculizumab and C5 deficiency on Escherichia coli ( E. coli )‐induced tissue factor (TF) <t>mRNA</t> levels and monocyte TF surface expression on whole blood monocytes. Whole blood from healthy donors was preincubated with phosphate‐buffered saline (PBS), eculizumab (Eculiz.) or control antibody (Ctrl.Ab.). The blood samples were either processed immediately (time zero samples) or incubated for 120 min at 37°C with PBS (control) or 1 × 10 7 /ml E. coli . TF mRNA expression (a,c) was examined using real‐time quantitative PCR <t>(qPCR)</t> and expressed as the relative quantity (RQ) compared to the PBS control (set to 1). Monocyte TF surface expression (b,d) was examined using flow cytometry and given as median flow intensity (MFI). Whole blood from the C5‐deficient individual was incubated with PBS, purified complement component C5 or human serum albumin (HSA) prior to incubation with E. coli (c,d). The results are given as means ± standard deviation (s.d.) for normal blood experiments ( n = 6), and as mean (line) and scatterplot of two experiments performed on two different days with C5D blood. * P
    Real Time Quantitative Polymerase Chain Reaction Qpcr, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 131 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher real time quantitative pcr qpcr total rna
    Pcf11 Expression Is Regulated by IPA (A) Schematic of mouse Pcf11 gene. Multiple PASs are indicated. EPA, internal exonic polyadenylation; IPA, intronic polyadenylation; TPA, 3′ terminal exon polyadenylation. 3′READS data of brain, heart, and testis are shown. (B) Features of intron 1 compared with other introns in the mouse genome. Each feature has ten bins, and the bin containing intron 1 of Pcf11 is highlighted in red. Distance to TSS is based on all IPA sites in PolyA_DB. (C) <t>RT-qPCR</t> analysis of IPA and TPA isoform expression in siPcf11 versus siCtrl cells. Error bars are SD on the basis of three replicates. (D) Top: schematic showing two sgRNAs used to remove a region flanking the IPA site. Bottom: validation of IPA site knockout (IPA Pcf11 -KO) by genomic DNA <t>PCR</t> using primers flanking the KO region. WT, wild-type 4T1 cells. (E) RT-qPCR analysis of Pcf11 isoform expression in WT and IPA Pcf11 -KO cells. Error bars are SD on the basis of three replicates. (F) Western blot analysis of PCF11 in WT and IPA Pcf11 -KO cells. (G) 3′UTR APA changes in IPA Pcf11 -KO cells. (H) IPA changes in IPA Pcf11 -KO cells. IPA isoforms (all combined) were compared with TPA isoforms (all combined) in analysis. (I) IPA distribution maps of activated IPA events in IPA Pcf11 -KO cells. (J) Left: schematic of PASs around the transcription start site (TSS); right: metagene analysis of PAS usage around the TSS in WT and IPA Pcf1 1 -KO cells. (K) Gene expression changes in IPA Pcf11 -KO cells for different groups of genes on the basis of IPA site frequency in PolyA_DB. ***p
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    Thermo Fisher quantitative reverse transcription polymerase chain reaction rt qpcr total rna
    Long-chain non-coding <t>RNA</t> metastasis-related lung adenocarcinoma transcript 1 (lncRNA MALAT1) is highly expressed in the plasma and hypoxic human pulmonary artery smooth muscle cells (hPASMCs) of patients with PAH. ( A ) Quantitative reverse transcription-PCR (qRT-PCR) detection of plasma lncRNA MALAT1 levels. ( B ) Receiver operating curve (ROC) for the diagnosis of PAH based on plasma lncRNA MALAT1 level. ( C ) Comparison of lncRNA MALAT1 levels in hypoxic and non-hypoxic hPASMC cells by <t>RT-qPCR</t>
    Quantitative Reverse Transcription Polymerase Chain Reaction Rt Qpcr Total Rna, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 88/100, based on 23 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Droplet digital PCR for oocyst genome quantification. a One-dimensional scatter plots showing 18S ddPCR assay on positive (syn18s standards from 7.2 × 10 4 to 0.72 copies/µL) and negative human blood (“HGD”) extracts. Clear demarcation between positive and negative partitions is shown. Uninfected human blood extract reaction was used to determine a universal positive/negative threshold set at 1853. b Quantification of syn18s standards using ddPCR in triplicate (black circles indicate the median of each run with error bars showing 95% CI) compared to the predicted qPCR value (grey line). c One-dimensional scatter plots showing 18S ddPCR assay on 14 oocyst-positive midguts, with positive and negative partitions. Uninfected mosquito midguts (“Neg”) were used to determine a universal positive/negative threshold set at 1853. d Quantification of genomes per oocyst for the14 microscopy-confirmed oocyst-positive midguts using 18S qPCR and 18S ddPCR. Box plots indicate the median and whiskers show the minimum and maximum responses. Groups compared using Wilcoxon matched-pairs signed rank test ( p = 0.43)

    Journal: Malaria Journal

    Article Title: Assessing Plasmodium falciparum transmission in mosquito-feeding assays using quantitative PCR

    doi: 10.1186/s12936-018-2382-6

    Figure Lengend Snippet: Droplet digital PCR for oocyst genome quantification. a One-dimensional scatter plots showing 18S ddPCR assay on positive (syn18s standards from 7.2 × 10 4 to 0.72 copies/µL) and negative human blood (“HGD”) extracts. Clear demarcation between positive and negative partitions is shown. Uninfected human blood extract reaction was used to determine a universal positive/negative threshold set at 1853. b Quantification of syn18s standards using ddPCR in triplicate (black circles indicate the median of each run with error bars showing 95% CI) compared to the predicted qPCR value (grey line). c One-dimensional scatter plots showing 18S ddPCR assay on 14 oocyst-positive midguts, with positive and negative partitions. Uninfected mosquito midguts (“Neg”) were used to determine a universal positive/negative threshold set at 1853. d Quantification of genomes per oocyst for the14 microscopy-confirmed oocyst-positive midguts using 18S qPCR and 18S ddPCR. Box plots indicate the median and whiskers show the minimum and maximum responses. Groups compared using Wilcoxon matched-pairs signed rank test ( p = 0.43)

    Article Snippet: The synthetic DNA standards (syn18S DNA) consisted of the 18S qPCR target cloned into a pMA-T plasmid backbone (Thermo Fisher Scientific, Australia) and serially diluted in uninfected whole blood nucleic acid extract to give final concentrations of 7.2 × 104 to 0.1 DNA copies/µL.

    Techniques: Digital PCR, Real-time Polymerase Chain Reaction, Microscopy

    Effect of the anti‐human C5 antibody eculizumab and C5 deficiency on Escherichia coli ( E. coli )‐induced tissue factor (TF) mRNA levels and monocyte TF surface expression on whole blood monocytes. Whole blood from healthy donors was preincubated with phosphate‐buffered saline (PBS), eculizumab (Eculiz.) or control antibody (Ctrl.Ab.). The blood samples were either processed immediately (time zero samples) or incubated for 120 min at 37°C with PBS (control) or 1 × 10 7 /ml E. coli . TF mRNA expression (a,c) was examined using real‐time quantitative PCR (qPCR) and expressed as the relative quantity (RQ) compared to the PBS control (set to 1). Monocyte TF surface expression (b,d) was examined using flow cytometry and given as median flow intensity (MFI). Whole blood from the C5‐deficient individual was incubated with PBS, purified complement component C5 or human serum albumin (HSA) prior to incubation with E. coli (c,d). The results are given as means ± standard deviation (s.d.) for normal blood experiments ( n = 6), and as mean (line) and scatterplot of two experiments performed on two different days with C5D blood. * P

    Journal: Clinical and Experimental Immunology

    Article Title: Complement component 5 does not interfere with physiological hemostasis but is essential for Escherichia coli‐induced coagulation accompanied by Toll‐like receptor 4

    doi: 10.1111/cei.13240

    Figure Lengend Snippet: Effect of the anti‐human C5 antibody eculizumab and C5 deficiency on Escherichia coli ( E. coli )‐induced tissue factor (TF) mRNA levels and monocyte TF surface expression on whole blood monocytes. Whole blood from healthy donors was preincubated with phosphate‐buffered saline (PBS), eculizumab (Eculiz.) or control antibody (Ctrl.Ab.). The blood samples were either processed immediately (time zero samples) or incubated for 120 min at 37°C with PBS (control) or 1 × 10 7 /ml E. coli . TF mRNA expression (a,c) was examined using real‐time quantitative PCR (qPCR) and expressed as the relative quantity (RQ) compared to the PBS control (set to 1). Monocyte TF surface expression (b,d) was examined using flow cytometry and given as median flow intensity (MFI). Whole blood from the C5‐deficient individual was incubated with PBS, purified complement component C5 or human serum albumin (HSA) prior to incubation with E. coli (c,d). The results are given as means ± standard deviation (s.d.) for normal blood experiments ( n = 6), and as mean (line) and scatterplot of two experiments performed on two different days with C5D blood. * P

    Article Snippet: Real‐time quantitative polymerase chain reaction (qPCR) of TF mRNA levels The MagMaxTM for stabilized Blood Tubes RNA Isolation kit from Thermo Fisher Scientific (Vilnius, Lithuania) was used to isolate the RNA.

    Techniques: Expressing, Incubation, Real-time Polymerase Chain Reaction, Flow Cytometry, Cytometry, Purification, Standard Deviation

    Pcf11 Expression Is Regulated by IPA (A) Schematic of mouse Pcf11 gene. Multiple PASs are indicated. EPA, internal exonic polyadenylation; IPA, intronic polyadenylation; TPA, 3′ terminal exon polyadenylation. 3′READS data of brain, heart, and testis are shown. (B) Features of intron 1 compared with other introns in the mouse genome. Each feature has ten bins, and the bin containing intron 1 of Pcf11 is highlighted in red. Distance to TSS is based on all IPA sites in PolyA_DB. (C) RT-qPCR analysis of IPA and TPA isoform expression in siPcf11 versus siCtrl cells. Error bars are SD on the basis of three replicates. (D) Top: schematic showing two sgRNAs used to remove a region flanking the IPA site. Bottom: validation of IPA site knockout (IPA Pcf11 -KO) by genomic DNA PCR using primers flanking the KO region. WT, wild-type 4T1 cells. (E) RT-qPCR analysis of Pcf11 isoform expression in WT and IPA Pcf11 -KO cells. Error bars are SD on the basis of three replicates. (F) Western blot analysis of PCF11 in WT and IPA Pcf11 -KO cells. (G) 3′UTR APA changes in IPA Pcf11 -KO cells. (H) IPA changes in IPA Pcf11 -KO cells. IPA isoforms (all combined) were compared with TPA isoforms (all combined) in analysis. (I) IPA distribution maps of activated IPA events in IPA Pcf11 -KO cells. (J) Left: schematic of PASs around the transcription start site (TSS); right: metagene analysis of PAS usage around the TSS in WT and IPA Pcf1 1 -KO cells. (K) Gene expression changes in IPA Pcf11 -KO cells for different groups of genes on the basis of IPA site frequency in PolyA_DB. ***p

    Journal: Cell reports

    Article Title: Regulation of Intronic Polyadenylation by PCF11 Impacts mRNA Expression of Long Genes

    doi: 10.1016/j.celrep.2019.02.049

    Figure Lengend Snippet: Pcf11 Expression Is Regulated by IPA (A) Schematic of mouse Pcf11 gene. Multiple PASs are indicated. EPA, internal exonic polyadenylation; IPA, intronic polyadenylation; TPA, 3′ terminal exon polyadenylation. 3′READS data of brain, heart, and testis are shown. (B) Features of intron 1 compared with other introns in the mouse genome. Each feature has ten bins, and the bin containing intron 1 of Pcf11 is highlighted in red. Distance to TSS is based on all IPA sites in PolyA_DB. (C) RT-qPCR analysis of IPA and TPA isoform expression in siPcf11 versus siCtrl cells. Error bars are SD on the basis of three replicates. (D) Top: schematic showing two sgRNAs used to remove a region flanking the IPA site. Bottom: validation of IPA site knockout (IPA Pcf11 -KO) by genomic DNA PCR using primers flanking the KO region. WT, wild-type 4T1 cells. (E) RT-qPCR analysis of Pcf11 isoform expression in WT and IPA Pcf11 -KO cells. Error bars are SD on the basis of three replicates. (F) Western blot analysis of PCF11 in WT and IPA Pcf11 -KO cells. (G) 3′UTR APA changes in IPA Pcf11 -KO cells. (H) IPA changes in IPA Pcf11 -KO cells. IPA isoforms (all combined) were compared with TPA isoforms (all combined) in analysis. (I) IPA distribution maps of activated IPA events in IPA Pcf11 -KO cells. (J) Left: schematic of PASs around the transcription start site (TSS); right: metagene analysis of PAS usage around the TSS in WT and IPA Pcf1 1 -KO cells. (K) Gene expression changes in IPA Pcf11 -KO cells for different groups of genes on the basis of IPA site frequency in PolyA_DB. ***p

    Article Snippet: Real-time quantitative PCR (qPCR) Total RNA was extracted using TRIzol (Thermo Fisher) and treated with Turbo DNase (Thermo Fisher). cDNA was synthesized using oligo(dT) and M-MLV reverse transcriptase with 1.5–2 μg of RNA. cDNA was then mixed with real-time PCR primers and Luna qPCR master mix (NEB).

    Techniques: Expressing, Indirect Immunoperoxidase Assay, Quantitative RT-PCR, Knock-Out, Polymerase Chain Reaction, Western Blot

    Long-chain non-coding RNA metastasis-related lung adenocarcinoma transcript 1 (lncRNA MALAT1) is highly expressed in the plasma and hypoxic human pulmonary artery smooth muscle cells (hPASMCs) of patients with PAH. ( A ) Quantitative reverse transcription-PCR (qRT-PCR) detection of plasma lncRNA MALAT1 levels. ( B ) Receiver operating curve (ROC) for the diagnosis of PAH based on plasma lncRNA MALAT1 level. ( C ) Comparison of lncRNA MALAT1 levels in hypoxic and non-hypoxic hPASMC cells by RT-qPCR

    Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

    Article Title: Long-Chain Non-Coding RNA Metastasis-Related Lung Adenocarcinoma Transcript 1 (MALAT1) Promotes the Proliferation and Migration of Human Pulmonary Artery Smooth Muscle Cells (hPASMCs) by Regulating the MicroRNA-503 (miR-503)/Toll-Like Receptor 4 (TLR4) Signal Axis

    doi: 10.12659/MSM.923123

    Figure Lengend Snippet: Long-chain non-coding RNA metastasis-related lung adenocarcinoma transcript 1 (lncRNA MALAT1) is highly expressed in the plasma and hypoxic human pulmonary artery smooth muscle cells (hPASMCs) of patients with PAH. ( A ) Quantitative reverse transcription-PCR (qRT-PCR) detection of plasma lncRNA MALAT1 levels. ( B ) Receiver operating curve (ROC) for the diagnosis of PAH based on plasma lncRNA MALAT1 level. ( C ) Comparison of lncRNA MALAT1 levels in hypoxic and non-hypoxic hPASMC cells by RT-qPCR

    Article Snippet: Quantitative reverse transcription- polymerase chain reaction (RT-qPCR) Total RNA was isolated from peripheral blood mononuclear cells (PBMCs), hPASMCs, and subject plasma using TRIzol reagent (Thermo Fisher Scientific, Shanghai, China).

    Techniques: Polymerase Chain Reaction, Quantitative RT-PCR