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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 44921 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 44921 article reviews
    Price from $9.99 to $1999.99
    7500 fast real time pcr system - by Bioz Stars, 2020-09
    99/100 stars
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    99
    Thermo Fisher abi 7500 fast real time pcr system
    Transcription and expression of HCV core-IFN-α2a recombinant viruses. (A) Identification of pFBD-H1 and pFBD-H2. M: 1Kb Plus DNA ladder; pFBD-H1 and pFBD-H2 samples were digested by BamHI and EcoRI. (B) Identification of pFBD-H3 and pFBD-H4. M: 1Kb Plus DNA ladder; pFBD-H3 and pFBD-H4 samples were digested by BamHI and EcoRI. (C) <t>RT-PCR</t> results of HCV core gene in recombination viruses infect cells. Total RNA was isolated from Sf9 infected with vAcH1, vAcH2, vAcH3, or vAcH4. cDNA was synthesized with SuperScript First Strand Synthesis kit (Invitrogen) with 0.5 to 1.0 μg RNA according to the manufacturer’s instructions. Quantitative RT-PCR reactions were carried out using SYBR Green PCR master mix reagents on an <t>ABI</t> 7500 Fast Real-Time PCR System (Applied Biosystems). (D) Expression of HCV core-IFN-α2a fusion protein in recombinant virus infected cells. M: protein marker. Cells were harvested at 72 h post-infection (hpi) and lysed in SDS-PAGE loading buffer. Twenty micrograms of total protein was separated by SDS-PAGE and subjected to Western blot assay.
    Abi 7500 Fast Real Time Pcr System, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 25156 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/abi 7500 fast real time pcr system/product/Thermo Fisher
    Average 99 stars, based on 25156 article reviews
    Price from $9.99 to $1999.99
    abi 7500 fast real time pcr system - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    92
    Thermo Fisher 7500 fast real time pcr machine
    Transcription and expression of HCV core-IFN-α2a recombinant viruses. (A) Identification of pFBD-H1 and pFBD-H2. M: 1Kb Plus DNA ladder; pFBD-H1 and pFBD-H2 samples were digested by BamHI and EcoRI. (B) Identification of pFBD-H3 and pFBD-H4. M: 1Kb Plus DNA ladder; pFBD-H3 and pFBD-H4 samples were digested by BamHI and EcoRI. (C) <t>RT-PCR</t> results of HCV core gene in recombination viruses infect cells. Total RNA was isolated from Sf9 infected with vAcH1, vAcH2, vAcH3, or vAcH4. cDNA was synthesized with SuperScript First Strand Synthesis kit (Invitrogen) with 0.5 to 1.0 μg RNA according to the manufacturer’s instructions. Quantitative RT-PCR reactions were carried out using SYBR Green PCR master mix reagents on an <t>ABI</t> 7500 Fast Real-Time PCR System (Applied Biosystems). (D) Expression of HCV core-IFN-α2a fusion protein in recombinant virus infected cells. M: protein marker. Cells were harvested at 72 h post-infection (hpi) and lysed in SDS-PAGE loading buffer. Twenty micrograms of total protein was separated by SDS-PAGE and subjected to Western blot assay.
    7500 Fast Real Time Pcr Machine, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 1015 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/7500 fast real time pcr machine/product/Thermo Fisher
    Average 92 stars, based on 1015 article reviews
    Price from $9.99 to $1999.99
    7500 fast real time pcr machine - by Bioz Stars, 2020-09
    92/100 stars
      Buy from Supplier

    93
    Thermo Fisher 7500 fast real time pcr instrument
    Transcription and expression of HCV core-IFN-α2a recombinant viruses. (A) Identification of pFBD-H1 and pFBD-H2. M: 1Kb Plus DNA ladder; pFBD-H1 and pFBD-H2 samples were digested by BamHI and EcoRI. (B) Identification of pFBD-H3 and pFBD-H4. M: 1Kb Plus DNA ladder; pFBD-H3 and pFBD-H4 samples were digested by BamHI and EcoRI. (C) <t>RT-PCR</t> results of HCV core gene in recombination viruses infect cells. Total RNA was isolated from Sf9 infected with vAcH1, vAcH2, vAcH3, or vAcH4. cDNA was synthesized with SuperScript First Strand Synthesis kit (Invitrogen) with 0.5 to 1.0 μg RNA according to the manufacturer’s instructions. Quantitative RT-PCR reactions were carried out using SYBR Green PCR master mix reagents on an <t>ABI</t> 7500 Fast Real-Time PCR System (Applied Biosystems). (D) Expression of HCV core-IFN-α2a fusion protein in recombinant virus infected cells. M: protein marker. Cells were harvested at 72 h post-infection (hpi) and lysed in SDS-PAGE loading buffer. Twenty micrograms of total protein was separated by SDS-PAGE and subjected to Western blot assay.
    7500 Fast Real Time Pcr Instrument, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 1089 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/7500 fast real time pcr instrument/product/Thermo Fisher
    Average 93 stars, based on 1089 article reviews
    Price from $9.99 to $1999.99
    7500 fast real time pcr instrument - by Bioz Stars, 2020-09
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      Buy from Supplier

    99
    Thermo Fisher 7500 fast dx real time pcr instrument
    Transcription and expression of HCV core-IFN-α2a recombinant viruses. (A) Identification of pFBD-H1 and pFBD-H2. M: 1Kb Plus DNA ladder; pFBD-H1 and pFBD-H2 samples were digested by BamHI and EcoRI. (B) Identification of pFBD-H3 and pFBD-H4. M: 1Kb Plus DNA ladder; pFBD-H3 and pFBD-H4 samples were digested by BamHI and EcoRI. (C) <t>RT-PCR</t> results of HCV core gene in recombination viruses infect cells. Total RNA was isolated from Sf9 infected with vAcH1, vAcH2, vAcH3, or vAcH4. cDNA was synthesized with SuperScript First Strand Synthesis kit (Invitrogen) with 0.5 to 1.0 μg RNA according to the manufacturer’s instructions. Quantitative RT-PCR reactions were carried out using SYBR Green PCR master mix reagents on an <t>ABI</t> 7500 Fast Real-Time PCR System (Applied Biosystems). (D) Expression of HCV core-IFN-α2a fusion protein in recombinant virus infected cells. M: protein marker. Cells were harvested at 72 h post-infection (hpi) and lysed in SDS-PAGE loading buffer. Twenty micrograms of total protein was separated by SDS-PAGE and subjected to Western blot assay.
    7500 Fast Dx Real Time Pcr Instrument, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 291 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/7500 fast dx real time pcr instrument/product/Thermo Fisher
    Average 99 stars, based on 291 article reviews
    Price from $9.99 to $1999.99
    7500 fast dx real time pcr instrument - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    91
    thermo fisher abi 7500 fast real time pcr instrument
    Transcription and expression of HCV core-IFN-α2a recombinant viruses. (A) Identification of pFBD-H1 and pFBD-H2. M: 1Kb Plus DNA ladder; pFBD-H1 and pFBD-H2 samples were digested by BamHI and EcoRI. (B) Identification of pFBD-H3 and pFBD-H4. M: 1Kb Plus DNA ladder; pFBD-H3 and pFBD-H4 samples were digested by BamHI and EcoRI. (C) <t>RT-PCR</t> results of HCV core gene in recombination viruses infect cells. Total RNA was isolated from Sf9 infected with vAcH1, vAcH2, vAcH3, or vAcH4. cDNA was synthesized with SuperScript First Strand Synthesis kit (Invitrogen) with 0.5 to 1.0 μg RNA according to the manufacturer’s instructions. Quantitative RT-PCR reactions were carried out using SYBR Green PCR master mix reagents on an <t>ABI</t> 7500 Fast Real-Time PCR System (Applied Biosystems). (D) Expression of HCV core-IFN-α2a fusion protein in recombinant virus infected cells. M: protein marker. Cells were harvested at 72 h post-infection (hpi) and lysed in SDS-PAGE loading buffer. Twenty micrograms of total protein was separated by SDS-PAGE and subjected to Western blot assay.
    Abi 7500 Fast Real Time Pcr Instrument, supplied by thermo fisher, used in various techniques. Bioz Stars score: 91/100, based on 360 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/abi 7500 fast real time pcr instrument/product/thermo fisher
    Average 91 stars, based on 360 article reviews
    Price from $9.99 to $1999.99
    abi 7500 fast real time pcr instrument - by Bioz Stars, 2020-09
    91/100 stars
      Buy from Supplier

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    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

    Transcription and expression of HCV core-IFN-α2a recombinant viruses. (A) Identification of pFBD-H1 and pFBD-H2. M: 1Kb Plus DNA ladder; pFBD-H1 and pFBD-H2 samples were digested by BamHI and EcoRI. (B) Identification of pFBD-H3 and pFBD-H4. M: 1Kb Plus DNA ladder; pFBD-H3 and pFBD-H4 samples were digested by BamHI and EcoRI. (C) RT-PCR results of HCV core gene in recombination viruses infect cells. Total RNA was isolated from Sf9 infected with vAcH1, vAcH2, vAcH3, or vAcH4. cDNA was synthesized with SuperScript First Strand Synthesis kit (Invitrogen) with 0.5 to 1.0 μg RNA according to the manufacturer’s instructions. Quantitative RT-PCR reactions were carried out using SYBR Green PCR master mix reagents on an ABI 7500 Fast Real-Time PCR System (Applied Biosystems). (D) Expression of HCV core-IFN-α2a fusion protein in recombinant virus infected cells. M: protein marker. Cells were harvested at 72 h post-infection (hpi) and lysed in SDS-PAGE loading buffer. Twenty micrograms of total protein was separated by SDS-PAGE and subjected to Western blot assay.

    Journal: Nanoscale Research Letters

    Article Title: Self-assembled HCV core virus-like particles targeted and inhibited tumor cell migration and invasion

    doi: 10.1186/1556-276X-8-401

    Figure Lengend Snippet: Transcription and expression of HCV core-IFN-α2a recombinant viruses. (A) Identification of pFBD-H1 and pFBD-H2. M: 1Kb Plus DNA ladder; pFBD-H1 and pFBD-H2 samples were digested by BamHI and EcoRI. (B) Identification of pFBD-H3 and pFBD-H4. M: 1Kb Plus DNA ladder; pFBD-H3 and pFBD-H4 samples were digested by BamHI and EcoRI. (C) RT-PCR results of HCV core gene in recombination viruses infect cells. Total RNA was isolated from Sf9 infected with vAcH1, vAcH2, vAcH3, or vAcH4. cDNA was synthesized with SuperScript First Strand Synthesis kit (Invitrogen) with 0.5 to 1.0 μg RNA according to the manufacturer’s instructions. Quantitative RT-PCR reactions were carried out using SYBR Green PCR master mix reagents on an ABI 7500 Fast Real-Time PCR System (Applied Biosystems). (D) Expression of HCV core-IFN-α2a fusion protein in recombinant virus infected cells. M: protein marker. Cells were harvested at 72 h post-infection (hpi) and lysed in SDS-PAGE loading buffer. Twenty micrograms of total protein was separated by SDS-PAGE and subjected to Western blot assay.

    Article Snippet: Quantitative RT-PCR reactions were carried out using SYBR Green PCR master mix reagents on an ABI 7500 Fast Real-Time PCR System (Applied Biosystems, Foster City, CA, USA).

    Techniques: Expressing, Recombinant, Reverse Transcription Polymerase Chain Reaction, Isolation, Infection, Synthesized, Quantitative RT-PCR, SYBR Green Assay, Polymerase Chain Reaction, Real-time Polymerase Chain Reaction, Marker, SDS Page, Western Blot

    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

    Diosgenin up-regulated DDX3 expression. A. mRNA level of DDX3 was measured by q-PCR in diosgenin treated HepG2 and SMMC-7721 cells using power SYBR Green PCR Master Mix on an ABI Prism 7500 Fast Real-Time PCR system. The relative expression of DDX3 was calculated. GAPDH was used as an endogenous reference. B. The expression of DDX3, Cyclin D1, p21, Notch-1, E-cadherin, and β-catenin was detected by Western blotting analysis in diosgenin treated HepG2 and SMMC-7721 cells. *P

    Journal: American Journal of Translational Research

    Article Title: Diosgenin increased DDX3 expression in hepatocellular carcinoma

    doi:

    Figure Lengend Snippet: Diosgenin up-regulated DDX3 expression. A. mRNA level of DDX3 was measured by q-PCR in diosgenin treated HepG2 and SMMC-7721 cells using power SYBR Green PCR Master Mix on an ABI Prism 7500 Fast Real-Time PCR system. The relative expression of DDX3 was calculated. GAPDH was used as an endogenous reference. B. The expression of DDX3, Cyclin D1, p21, Notch-1, E-cadherin, and β-catenin was detected by Western blotting analysis in diosgenin treated HepG2 and SMMC-7721 cells. *P

    Article Snippet: For the quantification of DDX3 mRNAs, 2 μg total RNA was reverse-transcribed into cDNA using RevertAid First Strand cDNA Synthesis Kit. qPCR was performed using Power SYBR Green PCR Master Mix on an ABI Prism 7500 Fast Real-Time PCR system (Applied Biosystems, Foster City, CA, USA), the relative expression of DDX3 was calculated.

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

    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

    Experimental verification of a 16-compartment digital assay with comparison to the performance of qPCR assays. ( A ) The graph shows the results for the same amount of reference DNA suspended in different elution buffers and quantified with conventional qPCR and with the synergistic PCR algorithm. Tests performed on Applied Biosystems 7500 Fast RT System on the IVD Cytomegalovirus PCR kit (GeneProof) according to the prescription. Elution buffers: (1) water, (2) AE elution buffer QIAamp DNA Mini Kit (Quiagen), (3) MBL5 NucleoMag Blood 200 uL (MACHEREY-NAGEL), (4) MagJET Whole Blood Genomic DNA Kit (Thermo Scientific). The gray line shows the expected distribution of results for Real-Time assay. ( B ) The graph shows the result of 24 runs of the synergistic assay, each on 16 partitions of the amplification mix. We conducted two series of 12 assays on the two elution buffers (1 and 4) that provided the largest difference in the result of the conventional qPCR analysis. The gray line shows the expected distribution of results from the synergistic assay used in the experiment, which should provide 60% precision of assessment. This distribution was also verified using 10,000 Monte-Carlo simulations.

    Journal: Scientific Reports

    Article Title: Calibration-free assays on standard real-time PCR devices

    doi: 10.1038/srep44854

    Figure Lengend Snippet: Experimental verification of a 16-compartment digital assay with comparison to the performance of qPCR assays. ( A ) The graph shows the results for the same amount of reference DNA suspended in different elution buffers and quantified with conventional qPCR and with the synergistic PCR algorithm. Tests performed on Applied Biosystems 7500 Fast RT System on the IVD Cytomegalovirus PCR kit (GeneProof) according to the prescription. Elution buffers: (1) water, (2) AE elution buffer QIAamp DNA Mini Kit (Quiagen), (3) MBL5 NucleoMag Blood 200 uL (MACHEREY-NAGEL), (4) MagJET Whole Blood Genomic DNA Kit (Thermo Scientific). The gray line shows the expected distribution of results for Real-Time assay. ( B ) The graph shows the result of 24 runs of the synergistic assay, each on 16 partitions of the amplification mix. We conducted two series of 12 assays on the two elution buffers (1 and 4) that provided the largest difference in the result of the conventional qPCR analysis. The gray line shows the expected distribution of results from the synergistic assay used in the experiment, which should provide 60% precision of assessment. This distribution was also verified using 10,000 Monte-Carlo simulations.

    Article Snippet: The PCR was conducted on the Applied Biosystems 7500 Fast Real-Time System according to the GeneProofs’ prescription.

    Techniques: Real-time Polymerase Chain Reaction, Polymerase Chain Reaction, Amplification

    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

    Effect of miR-196a on cell proliferation and apoptosis in vitro. (A) miR-196a expression was quantified by qRT–PCR in U87MG and T98G cells 48 h after transfection of miR-196a mimics, AMO-miR-196a, or negative control oligo. (B) Cell viability

    Journal: Neuro-Oncology

    Article Title: MiR-196a exerts its oncogenic effect in glioblastoma multiforme by inhibition of IκBα both in vitro and in vivo

    doi: 10.1093/neuonc/not307

    Figure Lengend Snippet: Effect of miR-196a on cell proliferation and apoptosis in vitro. (A) miR-196a expression was quantified by qRT–PCR in U87MG and T98G cells 48 h after transfection of miR-196a mimics, AMO-miR-196a, or negative control oligo. (B) Cell viability

    Article Snippet: Quantitative real-time polymerase chain reaction (qRT-PCR) was performed in triplicate in the ABI 7500HT fast real-time PCR System (Applied Biosystems) and normalized with U6 and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) endogenous control.

    Techniques: In Vitro, Expressing, Quantitative RT-PCR, Transfection, Negative Control

    Upregulation of USP5 and its role in in pancreatic cancer USP5 mRNA expression was quantified by realtime PCR (qRT-PCR) and normalized to ribosomal protein, large, P0 (RPLP0) mRNA levels. (A) Expression in malignant tissues (n=10) was significantly increased compared to healthy controls (n=8) or chronic pancreatitis tissues (n=6). CP = Chronic Pancreatitis. (B, C) Transient loss of USP5 function significantly reduced proliferation and viability of four different pancreatic cancer cell lines as assessed by their ability to metabolize MTT reagent (B, viability index) or incorporate BrdU agent (C, proliferation index) as compared to non-silencing control siRNA (siC) or untreated cells (NT). (D) USP5 knockdown however had no impact on the migration speed of PaTu-8988T cells, as assessed by Time Lapse Microscopy and automated cell tracking. si1, si2 and si3 = three specific siRNAs against USP5 . siC = non-silencing control. NT = untreated cells. *p

    Journal: Oncotarget

    Article Title: The deubiquitinating enzyme USP5 promotes pancreatic cancer via modulating cell cycle regulators

    doi: 10.18632/oncotarget.19882

    Figure Lengend Snippet: Upregulation of USP5 and its role in in pancreatic cancer USP5 mRNA expression was quantified by realtime PCR (qRT-PCR) and normalized to ribosomal protein, large, P0 (RPLP0) mRNA levels. (A) Expression in malignant tissues (n=10) was significantly increased compared to healthy controls (n=8) or chronic pancreatitis tissues (n=6). CP = Chronic Pancreatitis. (B, C) Transient loss of USP5 function significantly reduced proliferation and viability of four different pancreatic cancer cell lines as assessed by their ability to metabolize MTT reagent (B, viability index) or incorporate BrdU agent (C, proliferation index) as compared to non-silencing control siRNA (siC) or untreated cells (NT). (D) USP5 knockdown however had no impact on the migration speed of PaTu-8988T cells, as assessed by Time Lapse Microscopy and automated cell tracking. si1, si2 and si3 = three specific siRNAs against USP5 . siC = non-silencing control. NT = untreated cells. *p

    Article Snippet: Quantitative real time Reverse Transcription PCR (qRT-PCR) was performed using SYBR Green MasterMix (Applied Biosystems, USA) on a 7500 Fast Realtime PCR system (Applied Biosystems).

    Techniques: Expressing, Polymerase Chain Reaction, Quantitative RT-PCR, MTT Assay, Migration, Time-lapse Microscopy, Cell Tracking Assay