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  • 99
    Thermo Fisher quantitative real time pcr
    U1 small nuclear RNA (snRNA) selectively enriched in the insoluble fraction from Alzheimer’s disease (AD) brain homogenates A . Dot blot of 10 μg insoluble fraction from three controls and three AD cases. Ponceau S stain and 2,2,7-trimethylguanosine cap labeling is shown. B–F . Quantitative real-time <t>PCR</t> for U1, U2, U4, U5 and <t>U6</t> using cDNA reverse transcribed from control and AD insoluble fraction RNA. Average bar is show in each panel. 2^(-Delta delta Ct) values are shown. n = 4. *Statistical significance ( P = 0.003, Student’s t -test).
    Quantitative Real Time Pcr, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 40815 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/quantitative real time pcr/product/Thermo Fisher
    Average 99 stars, based on 40815 article reviews
    Price from $9.99 to $1999.99
    quantitative real time pcr - by Bioz Stars, 2020-05
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    99
    Thermo Fisher 7500 fast real time pcr system
    Chemical structure and DNA binding properties of PM01183. (A) Structure of PM01183; (B) binding to naked DNA. Drugs were incubated with a 250 bp <t>PCR</t> product at 25°C during 1 h and the electrophoresis was run in 2% agarose-TAE; (C) relative binding affinities for different DNA triplet sequences. PM01183 was incubated with the labelled double-strand oligodeoxynucleotides at 25°C during 1 h; then the melting assay was started in a <t>7500</t> Fast Real-Time PCR System by increasing the temperature up to 95°C in small steps of 1°C·min −1 . Analysis of the 1/C 50 parameter (drug affinity for specific DNA sequences) and ΔT m (max) (stability of the drug–DNA complex) were analysed with an in-house developed Visual Basic Application (VBA) running on Microsoft Excel. The values correspond to the mean and standard deviation from at least two independent experiments for each sequence.
    7500 Fast Real Time Pcr System, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 32854 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/7500 fast real time pcr system/product/Thermo Fisher
    Average 99 stars, based on 32854 article reviews
    Price from $9.99 to $1999.99
    7500 fast real time pcr system - by Bioz Stars, 2020-05
    99/100 stars
      Buy from Supplier

    Image Search Results


    U1 small nuclear RNA (snRNA) selectively enriched in the insoluble fraction from Alzheimer’s disease (AD) brain homogenates A . Dot blot of 10 μg insoluble fraction from three controls and three AD cases. Ponceau S stain and 2,2,7-trimethylguanosine cap labeling is shown. B–F . Quantitative real-time PCR for U1, U2, U4, U5 and U6 using cDNA reverse transcribed from control and AD insoluble fraction RNA. Average bar is show in each panel. 2^(-Delta delta Ct) values are shown. n = 4. *Statistical significance ( P = 0.003, Student’s t -test).

    Journal: Brain pathology (Zurich, Switzerland)

    Article Title: Aggregates of Small Nuclear Ribonucleic Acids (snRNAs) in Alzheimer’s Disease’

    doi: 10.1111/bpa.12133

    Figure Lengend Snippet: U1 small nuclear RNA (snRNA) selectively enriched in the insoluble fraction from Alzheimer’s disease (AD) brain homogenates A . Dot blot of 10 μg insoluble fraction from three controls and three AD cases. Ponceau S stain and 2,2,7-trimethylguanosine cap labeling is shown. B–F . Quantitative real-time PCR for U1, U2, U4, U5 and U6 using cDNA reverse transcribed from control and AD insoluble fraction RNA. Average bar is show in each panel. 2^(-Delta delta Ct) values are shown. n = 4. *Statistical significance ( P = 0.003, Student’s t -test).

    Article Snippet: TaqMan primers ( ) for U1, U2, U4, U5 and U6 along with 250 ng of cDNA template were used to perform quantitative real-time PCR (Applied Biosystems 7500 Fast Real-Time PCR System, Foster City, CA, USA).

    Techniques: Dot Blot, Staining, Labeling, Real-time Polymerase Chain Reaction

    STING N153S triggers perivascular pulmonary inflammation, myeloid cell expansion, and T cell cytopenia independently of IRF3. (A) Expression of ISGs in STING N153S, STING knockout ( Sting −/− ), and Trex1 −/− MEFs as well as STING N153S and Sting −/− spleens. Each data point represents a unique ISG as fold increase compared with a WT littermate control. Data were assembled using ISG expression values from RNA-sequencing. (B) qRT-PCR analysis of ISGs in WT, STING N153S, Sting −/− MEFs. mRNA expression is normalized to WT. (C) ISG expression in two healthy control (HC) and SAVI patient skin fibroblasts. Data represent a selection of ISGs assembled from RNA-sequencing. (D) qRT-PCR analysis of ISGs in skin fibroblasts. Data are from two independent experiments with two technical replicates per ISG, normalized to HC. (E) Hematoxylin and eosin images of lungs from Irf3 −/− and Irf3 −/− STING N153S mice (4–6 mo of age). Pulmonary vessels are indicated (v) with black arrows highlighting perivascular immune cell infiltration. Pulmonary vessels are indicated (v) with arrows. Bar, 100 µm. (F) Quantitation of perivascular lung lesions in STING N153S ( n = 7) and Irf3 −/− N153S ( n = 4) mice. Results in F represent the mean ± SEM of data collected and analyzed in two independent experiments. (G) ISG mRNA expression in Irf3 −/− and Irf3 −/− N153S MEFs normalized to gene expression in Irf3 −/− MEFs. Data represent the mean ± SEM from two independent experiments with n = 3 biological replicates per genotype. (H–J) Representative dot plots (H) and quantification (I and J) of splenic myeloid cell populations from WT, STING N153S, Irf3 −/− , and Irf3 −/− STING N153S mice. Data represent the mean of two independent experiments with n = 4 or 5 mice per genotype. (K and L) Percentage (K) and total number (L) of CD8 + splenocytes from WT, STING N153S, Irf3 −/− , and Irf3 −/− STING N153S mice. Data were pooled from two independent experiments with n = 4 or 5 mice per genotype. ns, not significant; *, P

    Journal: The Journal of Experimental Medicine

    Article Title: STING-associated vasculopathy develops independently of IRF3 in mice

    doi: 10.1084/jem.20171351

    Figure Lengend Snippet: STING N153S triggers perivascular pulmonary inflammation, myeloid cell expansion, and T cell cytopenia independently of IRF3. (A) Expression of ISGs in STING N153S, STING knockout ( Sting −/− ), and Trex1 −/− MEFs as well as STING N153S and Sting −/− spleens. Each data point represents a unique ISG as fold increase compared with a WT littermate control. Data were assembled using ISG expression values from RNA-sequencing. (B) qRT-PCR analysis of ISGs in WT, STING N153S, Sting −/− MEFs. mRNA expression is normalized to WT. (C) ISG expression in two healthy control (HC) and SAVI patient skin fibroblasts. Data represent a selection of ISGs assembled from RNA-sequencing. (D) qRT-PCR analysis of ISGs in skin fibroblasts. Data are from two independent experiments with two technical replicates per ISG, normalized to HC. (E) Hematoxylin and eosin images of lungs from Irf3 −/− and Irf3 −/− STING N153S mice (4–6 mo of age). Pulmonary vessels are indicated (v) with black arrows highlighting perivascular immune cell infiltration. Pulmonary vessels are indicated (v) with arrows. Bar, 100 µm. (F) Quantitation of perivascular lung lesions in STING N153S ( n = 7) and Irf3 −/− N153S ( n = 4) mice. Results in F represent the mean ± SEM of data collected and analyzed in two independent experiments. (G) ISG mRNA expression in Irf3 −/− and Irf3 −/− N153S MEFs normalized to gene expression in Irf3 −/− MEFs. Data represent the mean ± SEM from two independent experiments with n = 3 biological replicates per genotype. (H–J) Representative dot plots (H) and quantification (I and J) of splenic myeloid cell populations from WT, STING N153S, Irf3 −/− , and Irf3 −/− STING N153S mice. Data represent the mean of two independent experiments with n = 4 or 5 mice per genotype. (K and L) Percentage (K) and total number (L) of CD8 + splenocytes from WT, STING N153S, Irf3 −/− , and Irf3 −/− STING N153S mice. Data were pooled from two independent experiments with n = 4 or 5 mice per genotype. ns, not significant; *, P

    Article Snippet: RNA was quantified and quality confirmed using a NanoDrop ND-1000 spectrophotometer. cDNA was synthesized with iScript cDNA synthesis kit (Bio-Rad). iTaq Universal SYBR Green Supermix (Bio-Rad) and an ABI-7500 Fast Real-Time PCR system (Applied Biosystems) were used for qRT-PCR.

    Techniques: Expressing, Knock-Out, RNA Sequencing Assay, Quantitative RT-PCR, Selection, Mouse Assay, Quantitation Assay

    Quantitative mRNA analysis of cytokine expression in intestinal IELs. Total RNA was extracted from freshly isolated IELs of BALB/c and DO11.10 mice ( N = 5), and cDNA of IL-2, IL-4, IL-6, IL-10, IL-17, IFN- γ , TNF- α , and TGB- β was made using PureLink Micro-to-Midi Total RNA Purification System (Invitrogen, SP, Brazil,). Quantitative PCR analysis was performed with the ABI 7500 Fast Real-Time PCR system (Applied Biosystems), in four replicates, with the TaqMan Mastermix (Applied Biosystems). Samples were normalized to 18S rRNA, and an arbitrary value of 1 was given to control group (naïve mice) for the normalization, and the remaining samples were plotted relative to that value. IELs from BALB/c mice immunized by ip route show an increase in the expression of mRNA of cytokines IL-10, IL-2, IFN- γ , TGF- β , and TNF- α . IELs from DO11.10 mice fed with OVA and challenged by ip route showed increased expression of mRNA for cytokines IFN- γ , TGF- β , and TNF- α . The results are representative of three independent experiments.

    Journal: Clinical and Developmental Immunology

    Article Title: Phenotypical and Functional Analysis of Intraepithelial Lymphocytes from Small Intestine of Mice in Oral Tolerance

    doi: 10.1155/2012/208054

    Figure Lengend Snippet: Quantitative mRNA analysis of cytokine expression in intestinal IELs. Total RNA was extracted from freshly isolated IELs of BALB/c and DO11.10 mice ( N = 5), and cDNA of IL-2, IL-4, IL-6, IL-10, IL-17, IFN- γ , TNF- α , and TGB- β was made using PureLink Micro-to-Midi Total RNA Purification System (Invitrogen, SP, Brazil,). Quantitative PCR analysis was performed with the ABI 7500 Fast Real-Time PCR system (Applied Biosystems), in four replicates, with the TaqMan Mastermix (Applied Biosystems). Samples were normalized to 18S rRNA, and an arbitrary value of 1 was given to control group (naïve mice) for the normalization, and the remaining samples were plotted relative to that value. IELs from BALB/c mice immunized by ip route show an increase in the expression of mRNA of cytokines IL-10, IL-2, IFN- γ , TGF- β , and TNF- α . IELs from DO11.10 mice fed with OVA and challenged by ip route showed increased expression of mRNA for cytokines IFN- γ , TGF- β , and TNF- α . The results are representative of three independent experiments.

    Article Snippet: The cDNA was made using SuperScript III First-Strand Synthesis Supermix (Invitrogen) with random primers (Invitrogen) and analyzed for IL-2, IL-4, IL-10, IL-6, IL-17, IFN-γ , TGF-β , and TNF-α gene expression by real-time PCR assay using an 7500 Fast Real-Time PCR (Applied Biosystems, Foster City, CA) according to the manufacturer's instructions; 18S ribosomal RNA (rRNA) was used as an internal control.

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

    Chemical structure and DNA binding properties of PM01183. (A) Structure of PM01183; (B) binding to naked DNA. Drugs were incubated with a 250 bp PCR product at 25°C during 1 h and the electrophoresis was run in 2% agarose-TAE; (C) relative binding affinities for different DNA triplet sequences. PM01183 was incubated with the labelled double-strand oligodeoxynucleotides at 25°C during 1 h; then the melting assay was started in a 7500 Fast Real-Time PCR System by increasing the temperature up to 95°C in small steps of 1°C·min −1 . Analysis of the 1/C 50 parameter (drug affinity for specific DNA sequences) and ΔT m (max) (stability of the drug–DNA complex) were analysed with an in-house developed Visual Basic Application (VBA) running on Microsoft Excel. The values correspond to the mean and standard deviation from at least two independent experiments for each sequence.

    Journal: British Journal of Pharmacology

    Article Title: PM01183, a new DNA minor groove covalent binder with potent in vitro and in vivo anti-tumour activity

    doi: 10.1111/j.1476-5381.2010.00945.x

    Figure Lengend Snippet: Chemical structure and DNA binding properties of PM01183. (A) Structure of PM01183; (B) binding to naked DNA. Drugs were incubated with a 250 bp PCR product at 25°C during 1 h and the electrophoresis was run in 2% agarose-TAE; (C) relative binding affinities for different DNA triplet sequences. PM01183 was incubated with the labelled double-strand oligodeoxynucleotides at 25°C during 1 h; then the melting assay was started in a 7500 Fast Real-Time PCR System by increasing the temperature up to 95°C in small steps of 1°C·min −1 . Analysis of the 1/C 50 parameter (drug affinity for specific DNA sequences) and ΔT m (max) (stability of the drug–DNA complex) were analysed with an in-house developed Visual Basic Application (VBA) running on Microsoft Excel. The values correspond to the mean and standard deviation from at least two independent experiments for each sequence.

    Article Snippet: For the experiments, we followed the methodology previously described ( ; ) using a 7500 Fast Real-Time PCR System (ABI Prism, Applied Biosystems, Foster City, CA, USA).

    Techniques: Binding Assay, Incubation, Polymerase Chain Reaction, Electrophoresis, Real-time Polymerase Chain Reaction, Standard Deviation, Sequencing

    Generation and characterization of iPSCs from human dermal fibroblasts (HDFs). (A) Schematic diagram represents a protocol to generate iPSCs from HDFs by retroviral transduction. (B) Morphology of HDFs on day 1 post-transduction (i), non ESC-like colonies on day 8 (ii), ESC-like colonies on day 13 (iii), iPS colonies on iHFF plate (iv) and an iPS colony on Matrigel-coated plate (v), scale bar (i) and (iii) = 200 µm, (ii), (iv) and (v) = 500 µm. (C) RT-qPCR analysis of pluripotency- and fibroblast-associated genes in HDF, HDF-iPSC1 and Chula2.hES cells. (D) RT-PCR analysis of pluripotent genes, OCT4 and NANOG, of all the established iPS clones, HDF-iPSC2-6, as compared to Chula2.hES cells. (E) Representative immunofluorescent images of pluripotent transcription factors and cell surface antigens of HDF-iPSC1 as compared to Chula2.hES cells, scale bar = 100 µm.

    Journal: PLoS ONE

    Article Title: Dual Small-Molecule Targeting of SMAD Signaling Stimulates Human Induced Pluripotent Stem Cells toward Neural Lineages

    doi: 10.1371/journal.pone.0106952

    Figure Lengend Snippet: Generation and characterization of iPSCs from human dermal fibroblasts (HDFs). (A) Schematic diagram represents a protocol to generate iPSCs from HDFs by retroviral transduction. (B) Morphology of HDFs on day 1 post-transduction (i), non ESC-like colonies on day 8 (ii), ESC-like colonies on day 13 (iii), iPS colonies on iHFF plate (iv) and an iPS colony on Matrigel-coated plate (v), scale bar (i) and (iii) = 200 µm, (ii), (iv) and (v) = 500 µm. (C) RT-qPCR analysis of pluripotency- and fibroblast-associated genes in HDF, HDF-iPSC1 and Chula2.hES cells. (D) RT-PCR analysis of pluripotent genes, OCT4 and NANOG, of all the established iPS clones, HDF-iPSC2-6, as compared to Chula2.hES cells. (E) Representative immunofluorescent images of pluripotent transcription factors and cell surface antigens of HDF-iPSC1 as compared to Chula2.hES cells, scale bar = 100 µm.

    Article Snippet: Gene expression analysis by RT-qPCR Total RNA was prepared using TRIzol reagent (Invitrogen Corporation). cDNA was prepared using 2 µg of RNA and reverse transcribed using SuperScript III First-Strand Synthesis System and Oligo (dT) primers (Invitrogen Corporation). qPCR analysis was carried out on 7500 Fast Real-time PCR machine (Applied Biosystems, California) using Power SYBR Green PCR Master Mix (Applied Biosystems).

    Techniques: Transduction, Quantitative RT-PCR, Reverse Transcription Polymerase Chain Reaction, Clone Assay