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    Thermo Fisher real time pcr thermocycler
    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) <t>RT-qPCR</t> analysis of pluripotency- and fibroblast-associated genes in HDF, HDF-iPSC1 and Chula2.hES cells. (D) <t>RT-PCR</t> 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.
    Real Time Pcr Thermocycler, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 9853 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher real time pcr system
    Effect of miR-196a on cell proliferation and apoptosis in vitro. (A) miR-196a expression was quantified by <t>qRT–PCR</t> in U87MG and T98G cells 48 h after transfection of miR-196a mimics, AMO-miR-196a, or negative control oligo. (B) Cell viability
    Real Time Pcr System, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 35357 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/real time pcr system/product/Thermo Fisher
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    Thermo Fisher stepone plus real time pcr systems
    Effect of miR-196a on cell proliferation and apoptosis in vitro. (A) miR-196a expression was quantified by <t>qRT–PCR</t> in U87MG and T98G cells 48 h after transfection of miR-196a mimics, AMO-miR-196a, or negative control oligo. (B) Cell viability
    Stepone Plus Real Time Pcr Systems, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 300 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    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

    Characterization and transduction of retinal organoids derived from hiPSC. (A) Schematic of a 70-day optimized protocol of hiPSC differentiation toward retinal organoids. (B) CRX and BRN3B expression in qRT-PCR analysis. Mann–Whitney Student’s test relative to the expression on day 35 at least four biological replicates ( N = 3) were used for each samples ∗∗ p

    Journal: Frontiers in Neuroscience

    Article Title: Optogenetic Light Sensors in Human Retinal Organoids

    doi: 10.3389/fnins.2018.00789

    Figure Lengend Snippet: Characterization and transduction of retinal organoids derived from hiPSC. (A) Schematic of a 70-day optimized protocol of hiPSC differentiation toward retinal organoids. (B) CRX and BRN3B expression in qRT-PCR analysis. Mann–Whitney Student’s test relative to the expression on day 35 at least four biological replicates ( N = 3) were used for each samples ∗∗ p

    Article Snippet: Quantitative PCR (qPCR) reactions were performed using Taqman Array Fast plates and Taqman Gene expression master mix (Thermo Fisher Scientific) for CRX and BRN3B and S18 in an Applied Biosystems real-time PCR machine (7500 Fast System).

    Techniques: Transduction, Derivative Assay, Expressing, Quantitative RT-PCR, MANN-WHITNEY

    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

    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

    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

    E 2 prevented Cx43 repression and gap junction defects induced by miR-23a overexpression in NRVCs. (A) qRT-PCR assay revealed that E 2 treatment inhibited the expression of miR-23a in NRVCs. (B) E 2 treatment prevented the reduction of Cx43 protein level induced by miR-23a in NRVCs. (C) The size of gap junction per intercalated discs. (D) The representative electron micrographs of NRVCs. Up: Full image of intercalated disk including fasciae adherentes junctions, desmosome and gap junctions. Gap junctions were pointed by arrows in dashed boxes. The gap junctions are pointed by arrows. Down: magnified images of gap junctions from dashed boxes of above images. The gap junctions are pointed by arrows. n=3. * P

    Journal: International Journal of Biological Sciences

    Article Title: MicroRNA-23a Participates in Estrogen Deficiency Induced Gap Junction Remodeling of Rats by Targeting GJA1

    doi: 10.7150/ijbs.10930

    Figure Lengend Snippet: E 2 prevented Cx43 repression and gap junction defects induced by miR-23a overexpression in NRVCs. (A) qRT-PCR assay revealed that E 2 treatment inhibited the expression of miR-23a in NRVCs. (B) E 2 treatment prevented the reduction of Cx43 protein level induced by miR-23a in NRVCs. (C) The size of gap junction per intercalated discs. (D) The representative electron micrographs of NRVCs. Up: Full image of intercalated disk including fasciae adherentes junctions, desmosome and gap junctions. Gap junctions were pointed by arrows in dashed boxes. The gap junctions are pointed by arrows. Down: magnified images of gap junctions from dashed boxes of above images. The gap junctions are pointed by arrows. n=3. * P

    Article Snippet: Levels of microRNAs and mRNAs were quantified by qRT-PCR performed on 7500 Fast Real-Time PCR Systems (Applied Biosystems, USA) using SYBR Green PCR Master Mix (Applied Biosystems, USA).

    Techniques: Over Expression, Quantitative RT-PCR, Expressing

    Decreased thymic CD4 output was observed in young but not middle-aged donors. (A) TREC counts (left) and CD31 + percentages in naïve CD4 T cells (right) of young and middle-aged donors. PBMC were isolated from leukapheresis blood and CD4 T cells were further isolated via positive selection. TREC content was examined longitudinally by Taqman Real time PCR. CD31 expression was determined by flow cytometry analysis. The average of all time points were presented with the SD ( P ≤ 0.05). (B) TREC frequencies in CD4 T cells isolated from young (left panels) and middle-aged (right panels) donors are shown. (C) CD31 + CD45RA + CD4 T cells.

    Journal: Frontiers in Immunology

    Article Title: Homeostasis of lymphocytes and monocytes in frequent blood donors

    doi: 10.3389/fimmu.2012.00271

    Figure Lengend Snippet: Decreased thymic CD4 output was observed in young but not middle-aged donors. (A) TREC counts (left) and CD31 + percentages in naïve CD4 T cells (right) of young and middle-aged donors. PBMC were isolated from leukapheresis blood and CD4 T cells were further isolated via positive selection. TREC content was examined longitudinally by Taqman Real time PCR. CD31 expression was determined by flow cytometry analysis. The average of all time points were presented with the SD ( P ≤ 0.05). (B) TREC frequencies in CD4 T cells isolated from young (left panels) and middle-aged (right panels) donors are shown. (C) CD31 + CD45RA + CD4 T cells.

    Article Snippet: The frequency of the δRec-ψJα TREC created when the TCRD segment is excised from the TCR β gene locus was determined using Taqman real-time PCR 7500 Fast Real-Time PCR System (Applied Biosystems).

    Techniques: Isolation, Selection, Real-time Polymerase Chain Reaction, Expressing, Flow Cytometry, Cytometry

    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