steponeplus real time pcr system  (Thermo Fisher)


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    Structured Review

    Thermo Fisher steponeplus real time pcr system
    Steponeplus Real Time Pcr System, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 3203 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/steponeplus real time pcr system/product/Thermo Fisher
    Average 99 stars, based on 3203 article reviews
    Price from $9.99 to $1999.99
    steponeplus real time pcr system - by Bioz Stars, 2020-02
    99/100 stars

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    Related Articles

    Amplification:

    Article Title: Serological and Molecular Identification of Brucella spp. in Pigs from Cairo and Giza Governorates, Egypt
    Article Snippet: .. Amplification and real-time fluorescence detection was carried out on a StepOnePlus™ Real-Time PCR System (Applied Biosystems® , Germany). ..

    Article Title: Matrix mineralization controls gene expression in osteoblastic cells
    Article Snippet: For QRT-PCR the QuantiTect SYBR Green PCR Kit (Qiagen) in combination with the StepOnePlus Real-Time PCR System (Applied Biosystems, Darmstadt, Germany) was employed. .. Each target was amplified in a total reaction volume of 20 μl per well, containing 10 μl QuantiTect SYBR Green PCR Master Mix, 2 μl QuantiTect Primer, 2 μl cDNA and 6 μl RNase-free water.

    Article Title: Development of Molecular Methods for Rapid Differentiation of Mycoplasma gallisepticum Vaccine Strains from Field Isolates
    Article Snippet: The GC clamp increases the melting temperature and the size of the amplicon. .. Melt-MAMA tests and the melt analysis of a PCR assay were optimized on an Applied Biosystems StepOnePlus real-time PCR system with the StepOne software version 2.3 (Thermo Fisher Scientific, Waltham, MA, USA).

    Synthesized:

    Article Title: Expression Analysis of the PITX2 Gene and Associations between Its Polymorphisms and Body Size and Carcass Traits in Chickens
    Article Snippet: Primers were synthesized by Tsingke Biotech Co., Ltd. (Hangzhou, China). .. Real-time PCR was performed using a StepOnePlus Real-Time PCR System (Applied Biosystems, Foster City, CA, USA) with a 20 μL PCR mixture including 2 μL of cDNA, 10 μL of 2 × SYBR Premix Ex TaqII, 0.4 μL ROX Reference Dye (50×) 0.8 μL of forward primer, 0.8 μL of reverse primer and 6 μL of nuclease-free water.

    Article Title: Rapamycin-upregulated miR-29b promotes mTORC1-hyperactive cell growth in TSC2-deficient cells by downregulating tumor suppressor retinoic acid receptor β (RARβ)
    Article Snippet: Real-time PCR was performed using an Applied Biosystems StepOnePlus Real-time PCR system (Thermo Fisher Scientific, USA) with miRNA-specific primers or gene-specific primers (see ). .. To quantify the absolute amounts of miR-29b, a synthesized Caenorhabditis elegans microRNA mimic (Thermo Fisher Scientific, USA), cel-miR-39 was used as the spike-in control. miRNA was isolated from cells using the PureLink miRNA isolation kit (Thermo Fisher Scientific, USA) following the manufacturer’s protocol and cel-miR-39 (250 pmol) was added to each sample when cells were lysed.

    Article Title: RNA‐binding protein NONO promotes breast cancer proliferation by post‐transcriptional regulation of SKP2 and E2F8, et al. RNA‐binding protein NONO promotes breast cancer proliferation by post‐transcriptional regulation of SKP2 and E2F8
    Article Snippet: .. 2.8 Quantitative reverse transcription polymerase chain reaction RNA was extracted from cells using ISOGEN (Nippon Gene) and single‐stranded cDNA was synthesized using Super Script III reverse transcriptase (Invitrogen) with oligo dT or random primers. qRT‐PCR was carried out on StepOnePlus Real‐Time PCR System (Thermo Fisher Scientific) using KAPA SYBR FAST qPCR Kit (KAPA Biosystems) and sets of gene‐specific primers. ..

    Quantitative RT-PCR:

    Article Title: Knockout of beta‐2 microglobulin reduces stem cell‐induced immune rejection and enhances ischaemic hindlimb repair via exosome/miR‐24/Bim pathway, et al. Knockout of beta‐2 microglobulin reduces stem cell‐induced immune rejection and enhances ischaemic hindlimb repair via exosome/miR‐24/Bim pathway
    Article Snippet: .. The expression levels of miRNAs were analysed by qRT‐PCR using Takara SYBR Premix Ex Taq (Tli RNaseH Plus) in a StepOnePlus Real‐Time PCR system (Applied Biosystems). ..

    Article Title: Placental Mesenchymal Stem Cell-Derived Extracellular Vesicles Promote Myelin Regeneration in an Animal Model of Multiple Sclerosis
    Article Snippet: Sybr Green-based RT-qPCR was used to quantify the expression of OL differentiation markers oligodendrocyte-specific molecules (MOG), ectonucleotide pyrophosphatase/phosphodiesterase 6 (Enpp6) and myelin associated glycoprotein (MAG). .. Data were analyzed using the StepOnePlus Real-Time PCR System (Thermo Fisher Scientific) and processed by the V2.3 StepOne software.

    Article Title: Rapamycin-upregulated miR-29b promotes mTORC1-hyperactive cell growth in TSC2-deficient cells by downregulating tumor suppressor retinoic acid receptor β (RARβ)
    Article Snippet: Paragraph title: RNA isolation, miRNA isolation, and RT-qPCR ... Real-time PCR was performed using an Applied Biosystems StepOnePlus Real-time PCR system (Thermo Fisher Scientific, USA) with miRNA-specific primers or gene-specific primers (see ).

    Article Title: Elevated Intraocular Pressure Causes Abnormal Reactivity of Mouse Retinal Arterioles
    Article Snippet: .. Quantitative real-time RT-PCR (qPCR) reactions were performed on a StepOnePlus™ Real-Time PCR System (Applied Biosystems) using SYBR® Green JumpStart™ Taq ReadyMix™ (Sigma-Aldrich, Munich, Germany) and 20 ng cDNA. ..

    Article Title: RNA‐binding protein NONO promotes breast cancer proliferation by post‐transcriptional regulation of SKP2 and E2F8, et al. RNA‐binding protein NONO promotes breast cancer proliferation by post‐transcriptional regulation of SKP2 and E2F8
    Article Snippet: .. 2.8 Quantitative reverse transcription polymerase chain reaction RNA was extracted from cells using ISOGEN (Nippon Gene) and single‐stranded cDNA was synthesized using Super Script III reverse transcriptase (Invitrogen) with oligo dT or random primers. qRT‐PCR was carried out on StepOnePlus Real‐Time PCR System (Thermo Fisher Scientific) using KAPA SYBR FAST qPCR Kit (KAPA Biosystems) and sets of gene‐specific primers. ..

    Article Title: Matrix mineralization controls gene expression in osteoblastic cells
    Article Snippet: .. For QRT-PCR the QuantiTect SYBR Green PCR Kit (Qiagen) in combination with the StepOnePlus Real-Time PCR System (Applied Biosystems, Darmstadt, Germany) was employed. .. The following ten commercial primer sets QuantiTect Primer Assay (Qiagen) were used to detect targets of interest: HMCN1 (Hs_HMCN1_1_SG, Cat. no.: QT00011228), NID2 (Hs_NID2_1_SG, Cat. no.: QT00055258), ITGA2 (Hs_ITGA2_1_SG, Cat. no.: QT00086695), VCAN (Hs_VCAN_1_SG, Cat. no.: QT00064064), GAPDH (Hs_GAPDH_1_SG, Cat. no.: QT00079247) and ACTB (Hs_ACTB_1_SG, Cat. no.: QT00095431).

    Article Title: RNF12 catalyzes BRF1 ubiquitination and regulates RNA polymerase III–dependent transcription
    Article Snippet: Paragraph title: Real-time RT–PCR ... Real-time PCR was performed using SYBR premix EX Taq (TaKaRa) and analyzed with the StepOnePlus real-time PCR system (Thermo Fisher Scientific).

    Real-time Polymerase Chain Reaction:

    Article Title: Expression Analysis of the PITX2 Gene and Associations between Its Polymorphisms and Body Size and Carcass Traits in Chickens
    Article Snippet: .. Real-time PCR was performed using a StepOnePlus Real-Time PCR System (Applied Biosystems, Foster City, CA, USA) with a 20 μL PCR mixture including 2 μL of cDNA, 10 μL of 2 × SYBR Premix Ex TaqII, 0.4 μL ROX Reference Dye (50×) 0.8 μL of forward primer, 0.8 μL of reverse primer and 6 μL of nuclease-free water. .. The 2× SYBR Premix Ex TaqII, ROX Reference Dye (50×) and nuclease-free water were contained in the TB Green Premix Ex Taq TM II (Takara, Dalian, China).

    Article Title: Knockout of beta‐2 microglobulin reduces stem cell‐induced immune rejection and enhances ischaemic hindlimb repair via exosome/miR‐24/Bim pathway, et al. Knockout of beta‐2 microglobulin reduces stem cell‐induced immune rejection and enhances ischaemic hindlimb repair via exosome/miR‐24/Bim pathway
    Article Snippet: .. The expression levels of miRNAs were analysed by qRT‐PCR using Takara SYBR Premix Ex Taq (Tli RNaseH Plus) in a StepOnePlus Real‐Time PCR system (Applied Biosystems). ..

    Article Title: Placental Mesenchymal Stem Cell-Derived Extracellular Vesicles Promote Myelin Regeneration in an Animal Model of Multiple Sclerosis
    Article Snippet: .. Data were analyzed using the StepOnePlus Real-Time PCR System (Thermo Fisher Scientific) and processed by the V2.3 StepOne software. .. For quantification, the mRNA expression level of interested genes in each sample was normalized to the internal control, housekeeping gene Hsp90 and fold change in gene expression was calculated based on the delta-delta Ct method as previously described [ , ].

    Article Title: A Biochemical Approach to Identify Direct MicroRNA Targets
    Article Snippet: .. SYBR Green master mix (Applied Biosystems cat# 4309155) and SteponePlus real-time PCR system (Applied Biosystems) for real-time PCR. ..

    Article Title: Rapamycin-upregulated miR-29b promotes mTORC1-hyperactive cell growth in TSC2-deficient cells by downregulating tumor suppressor retinoic acid receptor β (RARβ)
    Article Snippet: .. Real-time PCR was performed using an Applied Biosystems StepOnePlus Real-time PCR system (Thermo Fisher Scientific, USA) with miRNA-specific primers or gene-specific primers (see ). .. Values represent the average of three independent experiments, normalized to β-actin for gene expression or miR-320a for miR-29b expression.

    Article Title: Elevated Intraocular Pressure Causes Abnormal Reactivity of Mouse Retinal Arterioles
    Article Snippet: .. Quantitative real-time RT-PCR (qPCR) reactions were performed on a StepOnePlus™ Real-Time PCR System (Applied Biosystems) using SYBR® Green JumpStart™ Taq ReadyMix™ (Sigma-Aldrich, Munich, Germany) and 20 ng cDNA. ..

    Article Title: Selective breeding modifies mef2ca mutant incomplete penetrance by tuning the opposing Notch pathway
    Article Snippet: .. The mef2ca b1086 mutant allele we exclusively use in this study and the KASP genotyping (LGC) protocol for this allele using a StepOnePlus Real-Time PCR System (Applied Biosystems) has been previously described [ , , ]. ..

    Article Title: RNA‐binding protein NONO promotes breast cancer proliferation by post‐transcriptional regulation of SKP2 and E2F8, et al. RNA‐binding protein NONO promotes breast cancer proliferation by post‐transcriptional regulation of SKP2 and E2F8
    Article Snippet: .. 2.8 Quantitative reverse transcription polymerase chain reaction RNA was extracted from cells using ISOGEN (Nippon Gene) and single‐stranded cDNA was synthesized using Super Script III reverse transcriptase (Invitrogen) with oligo dT or random primers. qRT‐PCR was carried out on StepOnePlus Real‐Time PCR System (Thermo Fisher Scientific) using KAPA SYBR FAST qPCR Kit (KAPA Biosystems) and sets of gene‐specific primers. ..

    Article Title: Serological and Molecular Identification of Brucella spp. in Pigs from Cairo and Giza Governorates, Egypt
    Article Snippet: .. Amplification and real-time fluorescence detection was carried out on a StepOnePlus™ Real-Time PCR System (Applied Biosystems® , Germany). ..

    Article Title: Lysolipid receptor cross-talk regulates lymphatic endothelial junctions in lymph nodes
    Article Snippet: .. Expression of mRNA was quantitated using PerfeCTa SYBR Green FastMix Reaction Mixes (Quanta Bioscience) and the StepOnePlus Real-Time PCR System (Applied Biosystems) with cDNA equivalent to 7.5 ng total RNA. .. Primers used for real-time PCR include the following (5′–3′): HPRT-Fw, TGA​CAC​TGG​CAA​AAC​AAT​GCA; HPRT-Rv, GGT​CCT​TTT​CAC​CAG​CAA​GCT; SPNS2-Fw, AAC​GTG​CTC​AAC​TAC​CTG​GAC; SPNS2 -Rv, ATG​AAC​ACT​GAC​TGC​AGC​AG; LPAR1-Fw, ACT​GTG​GTC​ATT​GTG​CTT​GG; LPAR1-Rv, ACA​GCA​CAC​GTC​TAG​AAG​TAA​C; FAM156A-Fw, TAT​GCT​GTT​GGG​AGG​GAA​GC; and FAM156A -Rv, GCA​GTA​TCG​ACA​TTC​ACA​TCG​G.

    Article Title: Matrix mineralization controls gene expression in osteoblastic cells
    Article Snippet: .. For QRT-PCR the QuantiTect SYBR Green PCR Kit (Qiagen) in combination with the StepOnePlus Real-Time PCR System (Applied Biosystems, Darmstadt, Germany) was employed. .. The following ten commercial primer sets QuantiTect Primer Assay (Qiagen) were used to detect targets of interest: HMCN1 (Hs_HMCN1_1_SG, Cat. no.: QT00011228), NID2 (Hs_NID2_1_SG, Cat. no.: QT00055258), ITGA2 (Hs_ITGA2_1_SG, Cat. no.: QT00086695), VCAN (Hs_VCAN_1_SG, Cat. no.: QT00064064), GAPDH (Hs_GAPDH_1_SG, Cat. no.: QT00079247) and ACTB (Hs_ACTB_1_SG, Cat. no.: QT00095431).

    Article Title: Development of Molecular Methods for Rapid Differentiation of Mycoplasma gallisepticum Vaccine Strains from Field Isolates
    Article Snippet: .. Melt-MAMA tests and the melt analysis of a PCR assay were optimized on an Applied Biosystems StepOnePlus real-time PCR system with the StepOne software version 2.3 (Thermo Fisher Scientific, Waltham, MA, USA). .. Primer melting temperature ( Tm ) and general suitability were calculated using NetPrimer (Premier Biosoft International, Palo Alto, CA).

    Article Title: Expression of LRIG1, a Negative Regulator of EGFR, Is Dynamically Altered during Different Stages of Gastric Carcinogenesis
    Article Snippet: .. Next, 1 μg of DNase-treated total RNA was used to synthesize cDNA using the ImProm-II Reverse Transcription System (Promega). cDNA was analyzed using Power SYBR Green Master Mix (Applied Biosystems, Waltham, MA) on a StepOnePlus Real-Time PCR System (Applied Biosystems), using the following primers: 5′-GTCACGGTCGCTGCTAACTC-3′ (forward) and 5′-TGGAAGGTGAGGCCCTCTAT-3′ (reverse) for LRIG1, 5′-ATGCCCGCATTAGCTCTTAG-3′ (forward) and 5′-GCAACTTCCCAAAATGTGCC-3′ (reverse) for EGFR, and 5′-AGAGCTACGAGCTGCCTGAC-3′ (forward) and 5′-AGCACTGTGTTGGCGTACAG-3′ (reverse) for actin. .. Two shRNA lentiviral vectors (in pGIPZ lentiviral shRNA vector) for LRIG1 (V2LHS_229246 and V2LHS_404471) were purchased from Open Biosystems (Huntsville, AL).

    Article Title: A combined computational and experimental approach reveals the structure of a C/EBPβ–Spi1 interaction required for IL1B gene transcription
    Article Snippet: .. 20-μl qPCRs containing 2- Maxima SYBR Green/ROX qPCR Master Mix (Thermo Scientific, K0223), 250 n m of primers, and 3 μl of precipitated DNA were set up in Fast 96-well reaction plates (Applied Biosystems, 4346907). qPCRs were carried out in a StepOnePlus Applied Biosystems real-time instrument (Thermo Fisher, 4376600). ..

    Article Title: RNF12 catalyzes BRF1 ubiquitination and regulates RNA polymerase III–dependent transcription
    Article Snippet: .. Real-time PCR was performed using SYBR premix EX Taq (TaKaRa) and analyzed with the StepOnePlus real-time PCR system (Thermo Fisher Scientific). .. The expression levels of the examined RNA were normalized to acidic ribosomal phosphoprotein P0.

    Incubation:

    Article Title: Expression Analysis of the PITX2 Gene and Associations between Its Polymorphisms and Body Size and Carcass Traits in Chickens
    Article Snippet: Real-time PCR was performed using a StepOnePlus Real-Time PCR System (Applied Biosystems, Foster City, CA, USA) with a 20 μL PCR mixture including 2 μL of cDNA, 10 μL of 2 × SYBR Premix Ex TaqII, 0.4 μL ROX Reference Dye (50×) 0.8 μL of forward primer, 0.8 μL of reverse primer and 6 μL of nuclease-free water. .. Reactions were incubated in a 96-well optical plate (Applied Biosystems, American) at 95 °C for 30 s, followed by 40 cycles of 95 °C for 5 s, and 60 °C for 30 s. Each sample was analyzed in triplicate.

    Article Title: Matrix mineralization controls gene expression in osteoblastic cells
    Article Snippet: For QRT-PCR the QuantiTect SYBR Green PCR Kit (Qiagen) in combination with the StepOnePlus Real-Time PCR System (Applied Biosystems, Darmstadt, Germany) was employed. .. DNA amplification was initiated through an incubation step of 15 minutes at 95°C, followed by 40 cycles of a 15 s denaturation step at 95 °C, a 30 seconds annealing step at 55 °C and a 30 seconds extension step at 72 °C.

    Expressing:

    Article Title: Expression Analysis of the PITX2 Gene and Associations between Its Polymorphisms and Body Size and Carcass Traits in Chickens
    Article Snippet: Paragraph title: 2.4. Relative Expression of PITX2 mRNA ... Real-time PCR was performed using a StepOnePlus Real-Time PCR System (Applied Biosystems, Foster City, CA, USA) with a 20 μL PCR mixture including 2 μL of cDNA, 10 μL of 2 × SYBR Premix Ex TaqII, 0.4 μL ROX Reference Dye (50×) 0.8 μL of forward primer, 0.8 μL of reverse primer and 6 μL of nuclease-free water.

    Article Title: Knockout of beta‐2 microglobulin reduces stem cell‐induced immune rejection and enhances ischaemic hindlimb repair via exosome/miR‐24/Bim pathway, et al. Knockout of beta‐2 microglobulin reduces stem cell‐induced immune rejection and enhances ischaemic hindlimb repair via exosome/miR‐24/Bim pathway
    Article Snippet: .. The expression levels of miRNAs were analysed by qRT‐PCR using Takara SYBR Premix Ex Taq (Tli RNaseH Plus) in a StepOnePlus Real‐Time PCR system (Applied Biosystems). ..

    Article Title: Placental Mesenchymal Stem Cell-Derived Extracellular Vesicles Promote Myelin Regeneration in an Animal Model of Multiple Sclerosis
    Article Snippet: Sybr Green-based RT-qPCR was used to quantify the expression of OL differentiation markers oligodendrocyte-specific molecules (MOG), ectonucleotide pyrophosphatase/phosphodiesterase 6 (Enpp6) and myelin associated glycoprotein (MAG). .. Data were analyzed using the StepOnePlus Real-Time PCR System (Thermo Fisher Scientific) and processed by the V2.3 StepOne software.

    Article Title: Rapamycin-upregulated miR-29b promotes mTORC1-hyperactive cell growth in TSC2-deficient cells by downregulating tumor suppressor retinoic acid receptor β (RARβ)
    Article Snippet: For gene expression, quantitative real-time PCR (qPCR) was performed using Taqman Universal Master Mix II, with UNG and for miRNA expression qPCR was performed using Taqman Fast Advanced Master mix (Thermo Fisher Scientific, USA). .. Real-time PCR was performed using an Applied Biosystems StepOnePlus Real-time PCR system (Thermo Fisher Scientific, USA) with miRNA-specific primers or gene-specific primers (see ).

    Article Title: Lysolipid receptor cross-talk regulates lymphatic endothelial junctions in lymph nodes
    Article Snippet: .. Expression of mRNA was quantitated using PerfeCTa SYBR Green FastMix Reaction Mixes (Quanta Bioscience) and the StepOnePlus Real-Time PCR System (Applied Biosystems) with cDNA equivalent to 7.5 ng total RNA. .. Primers used for real-time PCR include the following (5′–3′): HPRT-Fw, TGA​CAC​TGG​CAA​AAC​AAT​GCA; HPRT-Rv, GGT​CCT​TTT​CAC​CAG​CAA​GCT; SPNS2-Fw, AAC​GTG​CTC​AAC​TAC​CTG​GAC; SPNS2 -Rv, ATG​AAC​ACT​GAC​TGC​AGC​AG; LPAR1-Fw, ACT​GTG​GTC​ATT​GTG​CTT​GG; LPAR1-Rv, ACA​GCA​CAC​GTC​TAG​AAG​TAA​C; FAM156A-Fw, TAT​GCT​GTT​GGG​AGG​GAA​GC; and FAM156A -Rv, GCA​GTA​TCG​ACA​TTC​ACA​TCG​G.

    Article Title: RNF12 catalyzes BRF1 ubiquitination and regulates RNA polymerase III–dependent transcription
    Article Snippet: Real-time PCR was performed using SYBR premix EX Taq (TaKaRa) and analyzed with the StepOnePlus real-time PCR system (Thermo Fisher Scientific). .. The expression levels of the examined RNA were normalized to acidic ribosomal phosphoprotein P0.

    Transfection:

    Article Title: A Biochemical Approach to Identify Direct MicroRNA Targets
    Article Snippet: iScript cDNA synthesis kit (BIO-RAD, cat # 170–8896) for reverse transfection. .. SYBR Green master mix (Applied Biosystems cat# 4309155) and SteponePlus real-time PCR system (Applied Biosystems) for real-time PCR.

    Generated:

    Article Title: Elevated Intraocular Pressure Causes Abnormal Reactivity of Mouse Retinal Arterioles
    Article Snippet: RNA was isolated using peqGOLD TriFast™ (PEQLAB), and cDNA was generated with the High-Capacity cDNA Reverse Transcription Kit (Applied Biosystems, Darmstadt, Germany). .. Quantitative real-time RT-PCR (qPCR) reactions were performed on a StepOnePlus™ Real-Time PCR System (Applied Biosystems) using SYBR® Green JumpStart™ Taq ReadyMix™ (Sigma-Aldrich, Munich, Germany) and 20 ng cDNA.

    Polymerase Chain Reaction:

    Article Title: Expression Analysis of the PITX2 Gene and Associations between Its Polymorphisms and Body Size and Carcass Traits in Chickens
    Article Snippet: .. Real-time PCR was performed using a StepOnePlus Real-Time PCR System (Applied Biosystems, Foster City, CA, USA) with a 20 μL PCR mixture including 2 μL of cDNA, 10 μL of 2 × SYBR Premix Ex TaqII, 0.4 μL ROX Reference Dye (50×) 0.8 μL of forward primer, 0.8 μL of reverse primer and 6 μL of nuclease-free water. .. The 2× SYBR Premix Ex TaqII, ROX Reference Dye (50×) and nuclease-free water were contained in the TB Green Premix Ex Taq TM II (Takara, Dalian, China).

    Article Title: Serological and Molecular Identification of Brucella spp. in Pigs from Cairo and Giza Governorates, Egypt
    Article Snippet: Briefly, the PCR reaction was done in a 15 μL multiplex PCR mixture with 2× TaqMan™ Environmental master mix (Applied Biosystems® , Germany), 0.2 µM of each primer, 0.1 µM of each probe, and 5 μL of template DNA. .. Amplification and real-time fluorescence detection was carried out on a StepOnePlus™ Real-Time PCR System (Applied Biosystems® , Germany).

    Article Title: Matrix mineralization controls gene expression in osteoblastic cells
    Article Snippet: .. For QRT-PCR the QuantiTect SYBR Green PCR Kit (Qiagen) in combination with the StepOnePlus Real-Time PCR System (Applied Biosystems, Darmstadt, Germany) was employed. .. The following ten commercial primer sets QuantiTect Primer Assay (Qiagen) were used to detect targets of interest: HMCN1 (Hs_HMCN1_1_SG, Cat. no.: QT00011228), NID2 (Hs_NID2_1_SG, Cat. no.: QT00055258), ITGA2 (Hs_ITGA2_1_SG, Cat. no.: QT00086695), VCAN (Hs_VCAN_1_SG, Cat. no.: QT00064064), GAPDH (Hs_GAPDH_1_SG, Cat. no.: QT00079247) and ACTB (Hs_ACTB_1_SG, Cat. no.: QT00095431).

    Article Title: Development of Molecular Methods for Rapid Differentiation of Mycoplasma gallisepticum Vaccine Strains from Field Isolates
    Article Snippet: .. Melt-MAMA tests and the melt analysis of a PCR assay were optimized on an Applied Biosystems StepOnePlus real-time PCR system with the StepOne software version 2.3 (Thermo Fisher Scientific, Waltham, MA, USA). .. Primer melting temperature ( Tm ) and general suitability were calculated using NetPrimer (Premier Biosoft International, Palo Alto, CA).

    Binding Assay:

    Article Title: A combined computational and experimental approach reveals the structure of a C/EBPβ–Spi1 interaction required for IL1B gene transcription
    Article Snippet: 20-μl qPCRs containing 2- Maxima SYBR Green/ROX qPCR Master Mix (Thermo Scientific, K0223), 250 n m of primers, and 3 μl of precipitated DNA were set up in Fast 96-well reaction plates (Applied Biosystems, 4346907). qPCRs were carried out in a StepOnePlus Applied Biosystems real-time instrument (Thermo Fisher, 4376600). .. Final enrichment values were adjusted by subtraction of the nonspecific IgG antibody binding.

    Fluorescence:

    Article Title: Selective breeding modifies mef2ca mutant incomplete penetrance by tuning the opposing Notch pathway
    Article Snippet: The mef2ca b1086 mutant allele we exclusively use in this study and the KASP genotyping (LGC) protocol for this allele using a StepOnePlus Real-Time PCR System (Applied Biosystems) has been previously described [ , , ]. .. For genotyping dlx5a , heterozygotes and homozygous mutants were discriminated from wild types based on EGFP fluorescence.

    Article Title: Serological and Molecular Identification of Brucella spp. in Pigs from Cairo and Giza Governorates, Egypt
    Article Snippet: .. Amplification and real-time fluorescence detection was carried out on a StepOnePlus™ Real-Time PCR System (Applied Biosystems® , Germany). ..

    Magnetic Beads:

    Article Title: A combined computational and experimental approach reveals the structure of a C/EBPβ–Spi1 interaction required for IL1B gene transcription
    Article Snippet: The samples were precipitated using 25 μl of Magna ChIP protein A+G magnetic beads (EMD Millipore, 16-663) at 4 °C for 3 h and subsequently washed with the following solutions: once with low-salt buffer (0.1% SDS, 1% Triton X-100, 2 m m EDTA, 20 m m Tris-HCl, pH 8.1, 150 m m NaCl), once with high-salt buffer (0.1% SDS, 1% Triton X-100, 2 m m EDTA, 20 m m Tris-HCl, pH 8.1, 550 m m NaCl), once with LiCl wash buffer (0.25 m LiCl, 1% Igepal-CA630, 1% deoxycholic acid, 1 m m EDTA, 10 m m Tris, pH 8.1), and twice with TE buffer (10 m m Tris-HCl, 1 m m EDTA, pH 8.0). .. 20-μl qPCRs containing 2- Maxima SYBR Green/ROX qPCR Master Mix (Thermo Scientific, K0223), 250 n m of primers, and 3 μl of precipitated DNA were set up in Fast 96-well reaction plates (Applied Biosystems, 4346907). qPCRs were carried out in a StepOnePlus Applied Biosystems real-time instrument (Thermo Fisher, 4376600).

    Mutagenesis:

    Article Title: Selective breeding modifies mef2ca mutant incomplete penetrance by tuning the opposing Notch pathway
    Article Snippet: .. The mef2ca b1086 mutant allele we exclusively use in this study and the KASP genotyping (LGC) protocol for this allele using a StepOnePlus Real-Time PCR System (Applied Biosystems) has been previously described [ , , ]. ..

    Isolation:

    Article Title: Placental Mesenchymal Stem Cell-Derived Extracellular Vesicles Promote Myelin Regeneration in an Animal Model of Multiple Sclerosis
    Article Snippet: RNA isolation was performed using the RNeasy Mini Kit’s manufacturer’s protocol (QIAGEN, Germantown, MD, USA), and RNA concentration was measured with the NanoDrop 2000 Spectrophotometer (Thermo Fisher Scientific). .. Data were analyzed using the StepOnePlus Real-Time PCR System (Thermo Fisher Scientific) and processed by the V2.3 StepOne software.

    Article Title: Rapamycin-upregulated miR-29b promotes mTORC1-hyperactive cell growth in TSC2-deficient cells by downregulating tumor suppressor retinoic acid receptor β (RARβ)
    Article Snippet: Paragraph title: RNA isolation, miRNA isolation, and RT-qPCR ... Real-time PCR was performed using an Applied Biosystems StepOnePlus Real-time PCR system (Thermo Fisher Scientific, USA) with miRNA-specific primers or gene-specific primers (see ).

    Article Title: Elevated Intraocular Pressure Causes Abnormal Reactivity of Mouse Retinal Arterioles
    Article Snippet: RNA was isolated using peqGOLD TriFast™ (PEQLAB), and cDNA was generated with the High-Capacity cDNA Reverse Transcription Kit (Applied Biosystems, Darmstadt, Germany). .. Quantitative real-time RT-PCR (qPCR) reactions were performed on a StepOnePlus™ Real-Time PCR System (Applied Biosystems) using SYBR® Green JumpStart™ Taq ReadyMix™ (Sigma-Aldrich, Munich, Germany) and 20 ng cDNA.

    Article Title: Lysolipid receptor cross-talk regulates lymphatic endothelial junctions in lymph nodes
    Article Snippet: Paragraph title: RNA isolation and quantitative real-time PCR ... Expression of mRNA was quantitated using PerfeCTa SYBR Green FastMix Reaction Mixes (Quanta Bioscience) and the StepOnePlus Real-Time PCR System (Applied Biosystems) with cDNA equivalent to 7.5 ng total RNA.

    Article Title: RNF12 catalyzes BRF1 ubiquitination and regulates RNA polymerase III–dependent transcription
    Article Snippet: Total RNA was isolated using TRIzol (Invitrogen). .. Real-time PCR was performed using SYBR premix EX Taq (TaKaRa) and analyzed with the StepOnePlus real-time PCR system (Thermo Fisher Scientific).

    Multiplex Assay:

    Article Title: Serological and Molecular Identification of Brucella spp. in Pigs from Cairo and Giza Governorates, Egypt
    Article Snippet: Briefly, the PCR reaction was done in a 15 μL multiplex PCR mixture with 2× TaqMan™ Environmental master mix (Applied Biosystems® , Germany), 0.2 µM of each primer, 0.1 µM of each probe, and 5 μL of template DNA. .. Amplification and real-time fluorescence detection was carried out on a StepOnePlus™ Real-Time PCR System (Applied Biosystems® , Germany).

    Purification:

    Article Title: Lysolipid receptor cross-talk regulates lymphatic endothelial junctions in lymph nodes
    Article Snippet: Total RNA was isolated using TRI reagent (Zymo Research) and further purified with the Direct-zol RNA MicroPrep kit (Zymo Research), treated with DNase (30 U/µg total RNA, QIAGEN), and reverse transcribed using qScript XLT cDNA SuperMix (Quanta Bioscience). .. Expression of mRNA was quantitated using PerfeCTa SYBR Green FastMix Reaction Mixes (Quanta Bioscience) and the StepOnePlus Real-Time PCR System (Applied Biosystems) with cDNA equivalent to 7.5 ng total RNA.

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: RNA‐binding protein NONO promotes breast cancer proliferation by post‐transcriptional regulation of SKP2 and E2F8, et al. RNA‐binding protein NONO promotes breast cancer proliferation by post‐transcriptional regulation of SKP2 and E2F8
    Article Snippet: .. 2.8 Quantitative reverse transcription polymerase chain reaction RNA was extracted from cells using ISOGEN (Nippon Gene) and single‐stranded cDNA was synthesized using Super Script III reverse transcriptase (Invitrogen) with oligo dT or random primers. qRT‐PCR was carried out on StepOnePlus Real‐Time PCR System (Thermo Fisher Scientific) using KAPA SYBR FAST qPCR Kit (KAPA Biosystems) and sets of gene‐specific primers. ..

    Chloramphenicol Acetyltransferase Assay:

    Article Title: Elevated Intraocular Pressure Causes Abnormal Reactivity of Mouse Retinal Arterioles
    Article Snippet: Quantitative PCR Messenger RNA for the hypoxic markers, hypoxia-inducible factor 1alpha (HIF-1α ) and vascular endothelial growth factor-A (VEGF-A), for the prooxidant redox enzymes, nicotinamide adenine dinucleotide phosphate oxidases 1 and 2 (NOX1 and NOX2), for the antioxidant redox enzymes, catalase (CAT), heme oxygenase 1 (HO-1) and glutathione peroxidases 1 and 4 (GPx1 and GPx4), for the cytokines, tumor necrosis factor alpha (TNF-α ), interferon gamma (IFN-γ ), interleukin 1beta (IL-1β ), and interleukins 2 and 12 (IL-2 and IL-12), and for the nitric oxide synthase (NOS) isoforms, eNOS, iNOS, and nNOS, was quantified in the whole retinal explants as described before [ ]. .. Quantitative real-time RT-PCR (qPCR) reactions were performed on a StepOnePlus™ Real-Time PCR System (Applied Biosystems) using SYBR® Green JumpStart™ Taq ReadyMix™ (Sigma-Aldrich, Munich, Germany) and 20 ng cDNA.

    Chromatin Immunoprecipitation:

    Article Title: A combined computational and experimental approach reveals the structure of a C/EBPβ–Spi1 interaction required for IL1B gene transcription
    Article Snippet: Paragraph title: Chromatin immunoprecipitation ... 20-μl qPCRs containing 2- Maxima SYBR Green/ROX qPCR Master Mix (Thermo Scientific, K0223), 250 n m of primers, and 3 μl of precipitated DNA were set up in Fast 96-well reaction plates (Applied Biosystems, 4346907). qPCRs were carried out in a StepOnePlus Applied Biosystems real-time instrument (Thermo Fisher, 4376600).

    Software:

    Article Title: Placental Mesenchymal Stem Cell-Derived Extracellular Vesicles Promote Myelin Regeneration in an Animal Model of Multiple Sclerosis
    Article Snippet: .. Data were analyzed using the StepOnePlus Real-Time PCR System (Thermo Fisher Scientific) and processed by the V2.3 StepOne software. .. For quantification, the mRNA expression level of interested genes in each sample was normalized to the internal control, housekeeping gene Hsp90 and fold change in gene expression was calculated based on the delta-delta Ct method as previously described [ , ].

    Article Title: Development of Molecular Methods for Rapid Differentiation of Mycoplasma gallisepticum Vaccine Strains from Field Isolates
    Article Snippet: .. Melt-MAMA tests and the melt analysis of a PCR assay were optimized on an Applied Biosystems StepOnePlus real-time PCR system with the StepOne software version 2.3 (Thermo Fisher Scientific, Waltham, MA, USA). .. Primer melting temperature ( Tm ) and general suitability were calculated using NetPrimer (Premier Biosoft International, Palo Alto, CA).

    SYBR Green Assay:

    Article Title: A Biochemical Approach to Identify Direct MicroRNA Targets
    Article Snippet: .. SYBR Green master mix (Applied Biosystems cat# 4309155) and SteponePlus real-time PCR system (Applied Biosystems) for real-time PCR. ..

    Article Title: Elevated Intraocular Pressure Causes Abnormal Reactivity of Mouse Retinal Arterioles
    Article Snippet: .. Quantitative real-time RT-PCR (qPCR) reactions were performed on a StepOnePlus™ Real-Time PCR System (Applied Biosystems) using SYBR® Green JumpStart™ Taq ReadyMix™ (Sigma-Aldrich, Munich, Germany) and 20 ng cDNA. ..

    Article Title: Lysolipid receptor cross-talk regulates lymphatic endothelial junctions in lymph nodes
    Article Snippet: .. Expression of mRNA was quantitated using PerfeCTa SYBR Green FastMix Reaction Mixes (Quanta Bioscience) and the StepOnePlus Real-Time PCR System (Applied Biosystems) with cDNA equivalent to 7.5 ng total RNA. .. Primers used for real-time PCR include the following (5′–3′): HPRT-Fw, TGA​CAC​TGG​CAA​AAC​AAT​GCA; HPRT-Rv, GGT​CCT​TTT​CAC​CAG​CAA​GCT; SPNS2-Fw, AAC​GTG​CTC​AAC​TAC​CTG​GAC; SPNS2 -Rv, ATG​AAC​ACT​GAC​TGC​AGC​AG; LPAR1-Fw, ACT​GTG​GTC​ATT​GTG​CTT​GG; LPAR1-Rv, ACA​GCA​CAC​GTC​TAG​AAG​TAA​C; FAM156A-Fw, TAT​GCT​GTT​GGG​AGG​GAA​GC; and FAM156A -Rv, GCA​GTA​TCG​ACA​TTC​ACA​TCG​G.

    Article Title: Matrix mineralization controls gene expression in osteoblastic cells
    Article Snippet: .. For QRT-PCR the QuantiTect SYBR Green PCR Kit (Qiagen) in combination with the StepOnePlus Real-Time PCR System (Applied Biosystems, Darmstadt, Germany) was employed. .. The following ten commercial primer sets QuantiTect Primer Assay (Qiagen) were used to detect targets of interest: HMCN1 (Hs_HMCN1_1_SG, Cat. no.: QT00011228), NID2 (Hs_NID2_1_SG, Cat. no.: QT00055258), ITGA2 (Hs_ITGA2_1_SG, Cat. no.: QT00086695), VCAN (Hs_VCAN_1_SG, Cat. no.: QT00064064), GAPDH (Hs_GAPDH_1_SG, Cat. no.: QT00079247) and ACTB (Hs_ACTB_1_SG, Cat. no.: QT00095431).

    Article Title: Expression of LRIG1, a Negative Regulator of EGFR, Is Dynamically Altered during Different Stages of Gastric Carcinogenesis
    Article Snippet: .. Next, 1 μg of DNase-treated total RNA was used to synthesize cDNA using the ImProm-II Reverse Transcription System (Promega). cDNA was analyzed using Power SYBR Green Master Mix (Applied Biosystems, Waltham, MA) on a StepOnePlus Real-Time PCR System (Applied Biosystems), using the following primers: 5′-GTCACGGTCGCTGCTAACTC-3′ (forward) and 5′-TGGAAGGTGAGGCCCTCTAT-3′ (reverse) for LRIG1, 5′-ATGCCCGCATTAGCTCTTAG-3′ (forward) and 5′-GCAACTTCCCAAAATGTGCC-3′ (reverse) for EGFR, and 5′-AGAGCTACGAGCTGCCTGAC-3′ (forward) and 5′-AGCACTGTGTTGGCGTACAG-3′ (reverse) for actin. .. Two shRNA lentiviral vectors (in pGIPZ lentiviral shRNA vector) for LRIG1 (V2LHS_229246 and V2LHS_404471) were purchased from Open Biosystems (Huntsville, AL).

    RNA Extraction:

    Article Title: Expression of LRIG1, a Negative Regulator of EGFR, Is Dynamically Altered during Different Stages of Gastric Carcinogenesis
    Article Snippet: Paragraph title: RNA Extraction and Real-Time Quantitative PCR ... Next, 1 μg of DNase-treated total RNA was used to synthesize cDNA using the ImProm-II Reverse Transcription System (Promega). cDNA was analyzed using Power SYBR Green Master Mix (Applied Biosystems, Waltham, MA) on a StepOnePlus Real-Time PCR System (Applied Biosystems), using the following primers: 5′-GTCACGGTCGCTGCTAACTC-3′ (forward) and 5′-TGGAAGGTGAGGCCCTCTAT-3′ (reverse) for LRIG1, 5′-ATGCCCGCATTAGCTCTTAG-3′ (forward) and 5′-GCAACTTCCCAAAATGTGCC-3′ (reverse) for EGFR, and 5′-AGAGCTACGAGCTGCCTGAC-3′ (forward) and 5′-AGCACTGTGTTGGCGTACAG-3′ (reverse) for actin.

    Agarose Gel Electrophoresis:

    Article Title: Development of Molecular Methods for Rapid Differentiation of Mycoplasma gallisepticum Vaccine Strains from Field Isolates
    Article Snippet: The temperature shift can be easily detected in the presence of intercalating fluorescent dye on a real-time PCR platform (melt-MAMA), and the difference in the sizes of the amplicons can be observed in agarose gel electrophoresis (agarose-MAMA), which enable the differentiation of the SNP-specific genotypes. .. Melt-MAMA tests and the melt analysis of a PCR assay were optimized on an Applied Biosystems StepOnePlus real-time PCR system with the StepOne software version 2.3 (Thermo Fisher Scientific, Waltham, MA, USA).

    Spectrophotometry:

    Article Title: Knockout of beta‐2 microglobulin reduces stem cell‐induced immune rejection and enhances ischaemic hindlimb repair via exosome/miR‐24/Bim pathway, et al. Knockout of beta‐2 microglobulin reduces stem cell‐induced immune rejection and enhances ischaemic hindlimb repair via exosome/miR‐24/Bim pathway
    Article Snippet: RNA concentrations were measured using a spectrophotometer (NanoDrop). .. The expression levels of miRNAs were analysed by qRT‐PCR using Takara SYBR Premix Ex Taq (Tli RNaseH Plus) in a StepOnePlus Real‐Time PCR system (Applied Biosystems).

    Article Title: Placental Mesenchymal Stem Cell-Derived Extracellular Vesicles Promote Myelin Regeneration in an Animal Model of Multiple Sclerosis
    Article Snippet: RNA isolation was performed using the RNeasy Mini Kit’s manufacturer’s protocol (QIAGEN, Germantown, MD, USA), and RNA concentration was measured with the NanoDrop 2000 Spectrophotometer (Thermo Fisher Scientific). .. Data were analyzed using the StepOnePlus Real-Time PCR System (Thermo Fisher Scientific) and processed by the V2.3 StepOne software.

    Concentration Assay:

    Article Title: Placental Mesenchymal Stem Cell-Derived Extracellular Vesicles Promote Myelin Regeneration in an Animal Model of Multiple Sclerosis
    Article Snippet: RNA isolation was performed using the RNeasy Mini Kit’s manufacturer’s protocol (QIAGEN, Germantown, MD, USA), and RNA concentration was measured with the NanoDrop 2000 Spectrophotometer (Thermo Fisher Scientific). .. Data were analyzed using the StepOnePlus Real-Time PCR System (Thermo Fisher Scientific) and processed by the V2.3 StepOne software.

    Fluorescence In Situ Hybridization:

    Article Title: Selective breeding modifies mef2ca mutant incomplete penetrance by tuning the opposing Notch pathway
    Article Snippet: Zebrafish strains and husbandry All fish were maintained and staged according to established protocols [ , ]. .. The mef2ca b1086 mutant allele we exclusively use in this study and the KASP genotyping (LGC) protocol for this allele using a StepOnePlus Real-Time PCR System (Applied Biosystems) has been previously described [ , , ].

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  • 90
    Thermo Fisher real time pcr system
    Genotyping assay for the detection of the mutant allele (with the BRCA1 ex9-12del mutation) and the wild-type allele (exon 11) by real-time <t>PCR</t> and <t>TaqMan</t> ® probes. The AD plot shows 2 groups; green color samples have the allele with the deletion, samples in red color possess exon 11 or the wild-type allele (in this group all the samples are present, the 6 negative and the 4 positive ones). Samples in blue color are negative samples for the deletion, which have no fluorescence and are classified by the software as negatives. Negative controls are shown in black squares. Each sample was processed by duplicate. The axis values represent relative fluorescence (∆Rn) between both dyes (FAM ™ for the mutant allele and HEX ™ for the wild-type allele). PCR, polymerase chain reaction; AD, allelic discrimination.
    Real Time Pcr System, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 2976 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/real time pcr system/product/Thermo Fisher
    Average 90 stars, based on 2976 article reviews
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    real time pcr system - by Bioz Stars, 2020-02
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    90
    Thermo Fisher steponeplus real time system
    p21-/- attenuates the antiproliferative effects of AzaC (a) p21 mRNA is significantly upregulated in CD4+ and CD8+ Teff following treatment with AzaC. Teffs were isolated from the spleens of B6. Foxp3 GFP × B6.CAG DSRED and nTregs were isolated from B6. Foxp3 GFP . Cells were co-cultured at a 1:10 ratio of nTregs to Teffs for 2 days in the presence of anti-CD3/CD28 beads (bead:cell 1:1; Invitrogen) and Xcyte medium supplemented with L-glutamine (4 mM), penicillin (100 U/mL), streptomycin (100 μg/mL), and human recombinant IL-2 (hIL-2; 500 U/mL). The activated T cells were cultured with AzaC (1 μM) or PBS for an additional 2 days. Cells were sorted using FACS Aria II (BD) to isolate nTregs (CD4+DSRED-FOXP3GFP+), CD4+ Teffs (CD4+DSRed+FOXP3GFP-), and CD8+ Teffs (CD8+DSRed+FOXP3GFP-) prior to RNA extraction. QPCR was performed on the Applied Biosystems <t>StepOnePlus</t> Real-Time System using pre-designed TaqMan® Gene Expression Assays (18S RNA Mm03928990 and p21 Mm04205640). Relative fold changes in expression were determined using the ΔΔCT method. AzaC treatment resulted in a 3.4 fold increase of p21 expression in CD4+ Teffs (FACS sorted to remove AzaC converted Tregs) (AzaC vs. PBS p
    Steponeplus Real Time System, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 3228 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    79
    Thermo Fisher applied biosystemstm thermo fisher scientific steponeplustm real time pcr system
    p21-/- attenuates the antiproliferative effects of AzaC (a) p21 mRNA is significantly upregulated in CD4+ and CD8+ Teff following treatment with AzaC. Teffs were isolated from the spleens of B6. Foxp3 GFP × B6.CAG DSRED and nTregs were isolated from B6. Foxp3 GFP . Cells were co-cultured at a 1:10 ratio of nTregs to Teffs for 2 days in the presence of anti-CD3/CD28 beads (bead:cell 1:1; Invitrogen) and Xcyte medium supplemented with L-glutamine (4 mM), penicillin (100 U/mL), streptomycin (100 μg/mL), and human recombinant IL-2 (hIL-2; 500 U/mL). The activated T cells were cultured with AzaC (1 μM) or PBS for an additional 2 days. Cells were sorted using FACS Aria II (BD) to isolate nTregs (CD4+DSRED-FOXP3GFP+), CD4+ Teffs (CD4+DSRed+FOXP3GFP-), and CD8+ Teffs (CD8+DSRed+FOXP3GFP-) prior to RNA extraction. QPCR was performed on the Applied Biosystems <t>StepOnePlus</t> Real-Time System using pre-designed TaqMan® Gene Expression Assays (18S RNA Mm03928990 and p21 Mm04205640). Relative fold changes in expression were determined using the ΔΔCT method. AzaC treatment resulted in a 3.4 fold increase of p21 expression in CD4+ Teffs (FACS sorted to remove AzaC converted Tregs) (AzaC vs. PBS p
    Applied Biosystemstm Thermo Fisher Scientific Steponeplustm Real Time Pcr System, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 79/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Genotyping assay for the detection of the mutant allele (with the BRCA1 ex9-12del mutation) and the wild-type allele (exon 11) by real-time PCR and TaqMan ® probes. The AD plot shows 2 groups; green color samples have the allele with the deletion, samples in red color possess exon 11 or the wild-type allele (in this group all the samples are present, the 6 negative and the 4 positive ones). Samples in blue color are negative samples for the deletion, which have no fluorescence and are classified by the software as negatives. Negative controls are shown in black squares. Each sample was processed by duplicate. The axis values represent relative fluorescence (∆Rn) between both dyes (FAM ™ for the mutant allele and HEX ™ for the wild-type allele). PCR, polymerase chain reaction; AD, allelic discrimination.

    Journal: Molecular Medicine Reports

    Article Title: A novel method to detect the Mexican founder mutation BRCA1 ex9-12del associated with breast and ovarian cancer using quantitative polymerase chain reaction and TaqMan® probes

    doi: 10.3892/mmr.2018.9141

    Figure Lengend Snippet: Genotyping assay for the detection of the mutant allele (with the BRCA1 ex9-12del mutation) and the wild-type allele (exon 11) by real-time PCR and TaqMan ® probes. The AD plot shows 2 groups; green color samples have the allele with the deletion, samples in red color possess exon 11 or the wild-type allele (in this group all the samples are present, the 6 negative and the 4 positive ones). Samples in blue color are negative samples for the deletion, which have no fluorescence and are classified by the software as negatives. Negative controls are shown in black squares. Each sample was processed by duplicate. The axis values represent relative fluorescence (∆Rn) between both dyes (FAM ™ for the mutant allele and HEX ™ for the wild-type allele). PCR, polymerase chain reaction; AD, allelic discrimination.

    Article Snippet: qPCR with designed primers and TaqMan® probes Thermocycler StepOnePlus™ Real-Time PCR System (Thermo Fisher Scientific, Inc.) was used to perform the qPCR with TaqMan® probes.

    Techniques: Genotyping Assay, Mutagenesis, Real-time Polymerase Chain Reaction, Fluorescence, Software, Polymerase Chain Reaction

    Detection of the BRCA1 ex9-12del mutation by real-time PCR and TaqMan ® probe. Red amplification curves represent the fluorescent signal given by the FAM ™ dye from the TaqMan ® probe detecting the variant allele in the four positive samples, which had a mean Ct of 27.46. No amplification curve is observed for the negative samples neither for negative controls (horizontal red lines). The ‘Y’ axis values represent the fluorescent signal normalized of the FAM ™ dye with the ROX, whereas the ‘X’ axis represent number of cycles. PCR. PCR, polymerase chain reaction; ROX, passive reference dye.

    Journal: Molecular Medicine Reports

    Article Title: A novel method to detect the Mexican founder mutation BRCA1 ex9-12del associated with breast and ovarian cancer using quantitative polymerase chain reaction and TaqMan® probes

    doi: 10.3892/mmr.2018.9141

    Figure Lengend Snippet: Detection of the BRCA1 ex9-12del mutation by real-time PCR and TaqMan ® probe. Red amplification curves represent the fluorescent signal given by the FAM ™ dye from the TaqMan ® probe detecting the variant allele in the four positive samples, which had a mean Ct of 27.46. No amplification curve is observed for the negative samples neither for negative controls (horizontal red lines). The ‘Y’ axis values represent the fluorescent signal normalized of the FAM ™ dye with the ROX, whereas the ‘X’ axis represent number of cycles. PCR. PCR, polymerase chain reaction; ROX, passive reference dye.

    Article Snippet: qPCR with designed primers and TaqMan® probes Thermocycler StepOnePlus™ Real-Time PCR System (Thermo Fisher Scientific, Inc.) was used to perform the qPCR with TaqMan® probes.

    Techniques: Mutagenesis, Real-time Polymerase Chain Reaction, Amplification, Variant Assay, Polymerase Chain Reaction

    p21-/- attenuates the antiproliferative effects of AzaC (a) p21 mRNA is significantly upregulated in CD4+ and CD8+ Teff following treatment with AzaC. Teffs were isolated from the spleens of B6. Foxp3 GFP × B6.CAG DSRED and nTregs were isolated from B6. Foxp3 GFP . Cells were co-cultured at a 1:10 ratio of nTregs to Teffs for 2 days in the presence of anti-CD3/CD28 beads (bead:cell 1:1; Invitrogen) and Xcyte medium supplemented with L-glutamine (4 mM), penicillin (100 U/mL), streptomycin (100 μg/mL), and human recombinant IL-2 (hIL-2; 500 U/mL). The activated T cells were cultured with AzaC (1 μM) or PBS for an additional 2 days. Cells were sorted using FACS Aria II (BD) to isolate nTregs (CD4+DSRED-FOXP3GFP+), CD4+ Teffs (CD4+DSRed+FOXP3GFP-), and CD8+ Teffs (CD8+DSRed+FOXP3GFP-) prior to RNA extraction. QPCR was performed on the Applied Biosystems StepOnePlus Real-Time System using pre-designed TaqMan® Gene Expression Assays (18S RNA Mm03928990 and p21 Mm04205640). Relative fold changes in expression were determined using the ΔΔCT method. AzaC treatment resulted in a 3.4 fold increase of p21 expression in CD4+ Teffs (FACS sorted to remove AzaC converted Tregs) (AzaC vs. PBS p

    Journal: Journal of immunology (Baltimore, Md. : 1950)

    Article Title: Azacitidine mitigates GvHD via differential effects on the proliferation of T effectors and nTregs in vivo

    doi: 10.4049/jimmunol.1502399

    Figure Lengend Snippet: p21-/- attenuates the antiproliferative effects of AzaC (a) p21 mRNA is significantly upregulated in CD4+ and CD8+ Teff following treatment with AzaC. Teffs were isolated from the spleens of B6. Foxp3 GFP × B6.CAG DSRED and nTregs were isolated from B6. Foxp3 GFP . Cells were co-cultured at a 1:10 ratio of nTregs to Teffs for 2 days in the presence of anti-CD3/CD28 beads (bead:cell 1:1; Invitrogen) and Xcyte medium supplemented with L-glutamine (4 mM), penicillin (100 U/mL), streptomycin (100 μg/mL), and human recombinant IL-2 (hIL-2; 500 U/mL). The activated T cells were cultured with AzaC (1 μM) or PBS for an additional 2 days. Cells were sorted using FACS Aria II (BD) to isolate nTregs (CD4+DSRED-FOXP3GFP+), CD4+ Teffs (CD4+DSRed+FOXP3GFP-), and CD8+ Teffs (CD8+DSRed+FOXP3GFP-) prior to RNA extraction. QPCR was performed on the Applied Biosystems StepOnePlus Real-Time System using pre-designed TaqMan® Gene Expression Assays (18S RNA Mm03928990 and p21 Mm04205640). Relative fold changes in expression were determined using the ΔΔCT method. AzaC treatment resulted in a 3.4 fold increase of p21 expression in CD4+ Teffs (FACS sorted to remove AzaC converted Tregs) (AzaC vs. PBS p

    Article Snippet: QPCR was performed on the Applied Biosystems StepOnePlus Real-Time System (Thermo fisher) using pre-designed TaqMan® Gene Expression Assays (Life Technologies) (18S RNA Mm03928990 and p21 Mm04205640) according to manufacturer's instructions.

    Techniques: Isolation, Cell Culture, Recombinant, FACS, RNA Extraction, Real-time Polymerase Chain Reaction, Expressing