steponeplus thermocycler  (Thermo Fisher)


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    Structured Review

    Thermo Fisher steponeplus thermocycler
    Steponeplus Thermocycler, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 110 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/steponeplus thermocycler/product/Thermo Fisher
    Average 99 stars, based on 110 article reviews
    Price from $9.99 to $1999.99
    steponeplus thermocycler - by Bioz Stars, 2020-02
    99/100 stars

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    Related Articles

    Amplification:

    Article Title: L-Ferritin Binding to Scara5: A New Iron Traffic Pathway Potentially Implicated in Retinopathy
    Article Snippet: Before performing q-RT-PCR analysis, a final-point PCR was carried out for all genes to test the primers and verify the amplified products. .. Gene relative quantification was carried out using MicroAmp Optical 96-well plates (Applied Biosystems) in a StepOnePlus thermocycler (Applied Biosystems) in standard cycling conditions.

    Article Title: Deficient Vitamin E Uptake During Development Impairs Neural Tube Closure in Mice Lacking Lipoprotein Receptor SR-BI
    Article Snippet: .. The resulting cDNA was amplified by real time PCR with a StepOnePlus thermocycler (Applied Biosystems, CA) using the PowerUp SYBR Green master mix (Thermo, MA) and 100 nM of each primer. .. The primers, annealing temperatures and amplification efficiencies are listed in Supplementary Table .

    Article Title: Fibroblast Growth Factor Receptor Signaling Is Essential for Normal Mammary Gland Development and Stem Cell Function
    Article Snippet: RNA was collected through extraction with Trizol reagent (Invitrogen, Life Technologies, Carlsbad, CA) and cDNA templates were generated using a SuperScript III kit and 1 μ g RNA per sample according to the manufacturer’s protocol (Invitrogen, Carlsbad, CA). qPCR reactions were run using SYBR Green reagent (Applied Biosystems, Carlsbad, CA) on a StepOnePlus thermocycler (Applied Biosystems, Life Technologies, Carlsbad, CA) and fold changes were calculated using the Comparative CT (ΔΔCT) method and Step One software v2.0.1, Applied Biosystems, Life Technologies, Carlsbad, CA. .. Prior to analysis within sorted luminal and basal populations from the adult virgin mammary gland, RNA was amplified by the BCM Genomic and RNA Profiling Core using the NuGen WT-Ovation Kit (NuGen, San Carlos, CA).

    Article Title: Validation of reference genes for quantitative real-time PCR (qPCR) analysis of Actinobacillus suis
    Article Snippet: The amplification efficiencies of all primer pairs were between 95 and 102%. .. Real-time PCR was done in a StepOnePlus Thermocycler (Applied Biosystems, Foster City, CA) using a program with an initial denaturation step at 95°C for 30 seconds followed by 40 cycles of 95°C for 3 seconds and 60°C for 30 seconds.

    Article Title: Molecular assessment of trematode co-infection and intraspecific competition in molluscan intermediate hosts
    Article Snippet: A primer was deemed non-specific and discarded from further consideration if more than 10% of reactions with non-target DNA resulted in cycle thresholds (CT ) of 35 or lower or if post-cycling melt curve analysis indicated the presence of multiple amplification products. .. Real-time PCR and melt curve analyses were performed with Power SYBR Green MasterMix chemistry on a StepOnePlus thermocycler (Applied Biosystems).

    Article Title: In smokers, Sonic hedgehog modulates pulmonary endothelial function through vascular endothelial growth factor
    Article Snippet: .. After initial denaturation at 95 °C for 10 min, 20 ng of cDNA were amplified (using Gene Expression Master Mix, Life Technologies) in 40 annealing/extension cycles (95 °C for 15 s and 60 °C for 1 min) in a StepOnePlus thermocycler (Life Technologies). .. The sample’s fluorescence was measured after each cycle, and the threshold cycle (Ct) of the real-time PCR was defined as the point at which a fluorescence signal corresponding to the amplification of a PCR product was detectable.

    Article Title: Inhibition of HER2 Increases Jagged1-dependent Breast Cancer Stem Cells: Role for Membrane Jagged1
    Article Snippet: A melt curve was added after completion of the 40 cycles set by the StepOnePlus thermocycler manufacturer (Applied Biosystems, Foster City, CA). .. Ct value indicates the number of cycles that were required to detect the initiation of amplification of the cDNA amplicon.

    Synthesized:

    Article Title: Corticospinal-specific HCN expression in mouse motor cortex: Ih-dependent synaptic integration as a candidate microcircuit mechanism involved in motor control
    Article Snippet: RNA samples were treated with DNase I (Ambion, Austin, TX) to eliminate genomic DNA contamination, and cDNA was synthesized using qScript cDNA SuperMix (Quanta BioSciences, Gaithersburg, MD). .. Real-time PCR was performed using SYBR Green I on a StepOnePlus thermocycler (Applied Biosystems, Foster City, CA).

    Article Title: Strain-Specific Regulation of Striatal Phenotype in Drd2-eGFP BAC Transgenic Mice
    Article Snippet: Total RNA was isolated using RNeasy micro kit (Qiagen). cDNA was synthesized using qScript cDNA Supermix (Quanta Biosciences). .. Real-time PCR was performed using Fast SYBR Mastermix (Applied Biosystems) on a StepOnePlus thermocycler (Applied Biosystems).

    Article Title: Biosynthesis of organic photosensitizer Zn-porphyrin by diphtheria toxin repressor (DtxR)-mediated global upregulation of engineered heme biosynthesis pathway in Corynebacterium glutamicum
    Article Snippet: Total RNA with random primer was denatured at 65 °C for 10 minutes, and reverse-transcriptionally synthesized to cDNA templates by M-MLV reverse transcriptase. .. All oligonucleotides for qRT-PCR were designed based on target gene sequence (Supplementary Table ). qRT-PCR analysis was determined using a StepOnePlus thermocycler (Thermo Fisher Scientific, USA) with HiPi Real-Time PCR 2x Master Mix (SYBR green) according to the manufacturer’s instructions.

    Article Title: Local TNFR1 signaling licenses murine neutrophils for increased TLR-dependent cytokine and eicosanoid production
    Article Snippet: Neutrophils were lysed in RNAzol (MRC) and RNA was purified according to manufacturer’s instructions and concentrated using on a column (RNA Clean and Concentrator, Zymo Research). cDNA was prepared with iScript cDNA synthesis kit (Biorad), and quantitative PCR was performed with SYBR Green Master mix (Biorad) on a StepOnePlus thermocycler (Applied Biosystems). .. Primers were obtained from PrimerBank ( ) and synthesized by IDT (see ).

    Article Title: Inhibition of HER2 Increases Jagged1-dependent Breast Cancer Stem Cells: Role for Membrane Jagged1
    Article Snippet: After the cDNA was synthesized, real-time PCR was performed using iTaq™ SYBR® Green Supermix with ROX (BioRad, Hercules, CA) to detect the transcript levels of Notch receptors and Notch target genes. .. A melt curve was added after completion of the 40 cycles set by the StepOnePlus thermocycler manufacturer (Applied Biosystems, Foster City, CA).

    Quantitative RT-PCR:

    Article Title: Extracellular Vesicles Released by Leishmania (Leishmania) amazonensis Promote Disease Progression and Induce the Production of Different Cytokines in Macrophages and B-1 Cells
    Article Snippet: .. The real-time PCR reactions (qRT-PCR) were performed using the SYBR Green Real-Time PCR Master Mix (Applied Biosystems, Thermo Fisher Scientific) and the StepOnePlus thermocycler (Applied Biosystems) with equal amounts of each cDNA. ..

    Article Title: In smokers, Sonic hedgehog modulates pulmonary endothelial function through vascular endothelial growth factor
    Article Snippet: Paragraph title: RNA isolation and reverse transcriptase – quantitative polymerase chain reaction (RT-qPCR) analysis ... After initial denaturation at 95 °C for 10 min, 20 ng of cDNA were amplified (using Gene Expression Master Mix, Life Technologies) in 40 annealing/extension cycles (95 °C for 15 s and 60 °C for 1 min) in a StepOnePlus thermocycler (Life Technologies).

    Article Title: Biosynthesis of organic photosensitizer Zn-porphyrin by diphtheria toxin repressor (DtxR)-mediated global upregulation of engineered heme biosynthesis pathway in Corynebacterium glutamicum
    Article Snippet: .. All oligonucleotides for qRT-PCR were designed based on target gene sequence (Supplementary Table ). qRT-PCR analysis was determined using a StepOnePlus thermocycler (Thermo Fisher Scientific, USA) with HiPi Real-Time PCR 2x Master Mix (SYBR green) according to the manufacturer’s instructions. ..

    Article Title: Local TNFR1 signaling licenses murine neutrophils for increased TLR-dependent cytokine and eicosanoid production
    Article Snippet: Paragraph title: Quantitative RT-PCR ... Neutrophils were lysed in RNAzol (MRC) and RNA was purified according to manufacturer’s instructions and concentrated using on a column (RNA Clean and Concentrator, Zymo Research). cDNA was prepared with iScript cDNA synthesis kit (Biorad), and quantitative PCR was performed with SYBR Green Master mix (Biorad) on a StepOnePlus thermocycler (Applied Biosystems).

    Real-time Polymerase Chain Reaction:

    Article Title: L-Ferritin Binding to Scara5: A New Iron Traffic Pathway Potentially Implicated in Retinopathy
    Article Snippet: Paragraph title: Quantitative real-time PCR analysis of gene expression ... Gene relative quantification was carried out using MicroAmp Optical 96-well plates (Applied Biosystems) in a StepOnePlus thermocycler (Applied Biosystems) in standard cycling conditions.

    Article Title: Corticospinal-specific HCN expression in mouse motor cortex: Ih-dependent synaptic integration as a candidate microcircuit mechanism involved in motor control
    Article Snippet: .. Real-time PCR was performed using SYBR Green I on a StepOnePlus thermocycler (Applied Biosystems, Foster City, CA). .. The thermal cycling conditions were comprised of an initial denaturing step at 95°C for 3 min and 40 cycles at 95°C for 15 s, 60°C for 60 s, and 72°C for 30 s. The PCR cycle threshold values were measured within the exponential phase of the PCR reaction using StepOnePlus software (version 2.1, Applied Biosystems).

    Article Title: Modeling appendicular skeletal cartilage development with modified high-density micromass cultures of adult human bone marrow-derived mesenchymal progenitor cells
    Article Snippet: .. Quantitative real-time PCR was performed using a StepOnePlus thermocycler (Applied Biosystems) and SYBR Green Reaction Mix (Applied Biosystems). .. Expression of aggrecan (ACAN ) and collagen type II (COL2A1 ) was quantified to analyze chondrogenic differentiation.

    Article Title: Deficient Vitamin E Uptake During Development Impairs Neural Tube Closure in Mice Lacking Lipoprotein Receptor SR-BI
    Article Snippet: .. The resulting cDNA was amplified by real time PCR with a StepOnePlus thermocycler (Applied Biosystems, CA) using the PowerUp SYBR Green master mix (Thermo, MA) and 100 nM of each primer. .. The primers, annealing temperatures and amplification efficiencies are listed in Supplementary Table .

    Article Title: Fibroblast Growth Factor Receptor Signaling Is Essential for Normal Mammary Gland Development and Stem Cell Function
    Article Snippet: .. RNA was collected through extraction with Trizol reagent (Invitrogen, Life Technologies, Carlsbad, CA) and cDNA templates were generated using a SuperScript III kit and 1 μ g RNA per sample according to the manufacturer’s protocol (Invitrogen, Carlsbad, CA). qPCR reactions were run using SYBR Green reagent (Applied Biosystems, Carlsbad, CA) on a StepOnePlus thermocycler (Applied Biosystems, Life Technologies, Carlsbad, CA) and fold changes were calculated using the Comparative CT (ΔΔCT) method and Step One software v2.0.1, Applied Biosystems, Life Technologies, Carlsbad, CA. ..

    Article Title: Control of mitotic chromosome condensation by the fission yeast transcription factor Zas1
    Article Snippet: .. Transcript abundance was measured by qPCR using SYBR Green Mix (Thermo Fisher) on a StepOnePlus thermocycler (Applied Biosystems) using primer pairs described in Table S2, and for act1 as described previously ( ). .. For each qPCR run, primer efficiency was determined by RT reaction serial dilution.

    Article Title: Extracellular Vesicles Released by Leishmania (Leishmania) amazonensis Promote Disease Progression and Induce the Production of Different Cytokines in Macrophages and B-1 Cells
    Article Snippet: .. The real-time PCR reactions (qRT-PCR) were performed using the SYBR Green Real-Time PCR Master Mix (Applied Biosystems, Thermo Fisher Scientific) and the StepOnePlus thermocycler (Applied Biosystems) with equal amounts of each cDNA. ..

    Article Title: Strain-Specific Regulation of Striatal Phenotype in Drd2-eGFP BAC Transgenic Mice
    Article Snippet: .. Real-time PCR was performed using Fast SYBR Mastermix (Applied Biosystems) on a StepOnePlus thermocycler (Applied Biosystems). .. The thermal cycling conditions comprised of an initial denaturing step at 95°C for 20 s, and 40 cycles at 95°C for 3 s, 60°C for 30 s. The PCR cycle threshold (Ct) values were measured within the exponential phase of the PCR reaction using StepOnePlus software version 2.1 (Applied Biosystems).

    Article Title: Validation of reference genes for quantitative real-time PCR (qPCR) analysis of Actinobacillus suis
    Article Snippet: .. Real-time PCR was done in a StepOnePlus Thermocycler (Applied Biosystems, Foster City, CA) using a program with an initial denaturation step at 95°C for 30 seconds followed by 40 cycles of 95°C for 3 seconds and 60°C for 30 seconds. ..

    Article Title: Molecular assessment of trematode co-infection and intraspecific competition in molluscan intermediate hosts
    Article Snippet: .. Real-time PCR and melt curve analyses were performed with Power SYBR Green MasterMix chemistry on a StepOnePlus thermocycler (Applied Biosystems). .. Following an initial 10-minute denaturation and Taq activation at 95°C, reactions were cycled as follows, using a “stepdown” scheme to encourage reaction stringency: 2 cycles of 15s at 95°C and 60s at 60°C, 2 cycles of 15s at 95°C and 60s at 59°C, and 40 cycles of 15s at 95°C and 60s at 58°C.

    Article Title: In smokers, Sonic hedgehog modulates pulmonary endothelial function through vascular endothelial growth factor
    Article Snippet: Paragraph title: RNA isolation and reverse transcriptase – quantitative polymerase chain reaction (RT-qPCR) analysis ... After initial denaturation at 95 °C for 10 min, 20 ng of cDNA were amplified (using Gene Expression Master Mix, Life Technologies) in 40 annealing/extension cycles (95 °C for 15 s and 60 °C for 1 min) in a StepOnePlus thermocycler (Life Technologies).

    Article Title: Biosynthesis of organic photosensitizer Zn-porphyrin by diphtheria toxin repressor (DtxR)-mediated global upregulation of engineered heme biosynthesis pathway in Corynebacterium glutamicum
    Article Snippet: .. All oligonucleotides for qRT-PCR were designed based on target gene sequence (Supplementary Table ). qRT-PCR analysis was determined using a StepOnePlus thermocycler (Thermo Fisher Scientific, USA) with HiPi Real-Time PCR 2x Master Mix (SYBR green) according to the manufacturer’s instructions. ..

    Article Title: Local TNFR1 signaling licenses murine neutrophils for increased TLR-dependent cytokine and eicosanoid production
    Article Snippet: .. Neutrophils were lysed in RNAzol (MRC) and RNA was purified according to manufacturer’s instructions and concentrated using on a column (RNA Clean and Concentrator, Zymo Research). cDNA was prepared with iScript cDNA synthesis kit (Biorad), and quantitative PCR was performed with SYBR Green Master mix (Biorad) on a StepOnePlus thermocycler (Applied Biosystems). .. Primers were obtained from PrimerBank ( ) and synthesized by IDT (see ).

    Article Title: Inhibition of HER2 Increases Jagged1-dependent Breast Cancer Stem Cells: Role for Membrane Jagged1
    Article Snippet: Paragraph title: Real-time PCR ... A melt curve was added after completion of the 40 cycles set by the StepOnePlus thermocycler manufacturer (Applied Biosystems, Foster City, CA).

    Article Title: Development of Novel Single-Chain Antibodies against the Hydrophobic HPV-16 E5 Protein
    Article Snippet: .. The reaction was performed with Evagreen qPCR mastermix-Rox (ABM, Richmond, BC, Canada), following the manufacturer's instructions in a StepOnePlus thermocycler (Applied Biosystems). ..

    Modification:

    Article Title: Biosynthesis of organic photosensitizer Zn-porphyrin by diphtheria toxin repressor (DtxR)-mediated global upregulation of engineered heme biosynthesis pathway in Corynebacterium glutamicum
    Article Snippet: RNA Preparation and qRT-PCR To prepare total RNA, the CGPB02 and CGPB04 strains were cultivated in modified CGXII medium and harvested at 12 h. Total RNA in the cell pellet was extracted using a Hybrid-R prep kit (GeneAll, Korea) according to the manufacturer’s instructions. .. All oligonucleotides for qRT-PCR were designed based on target gene sequence (Supplementary Table ). qRT-PCR analysis was determined using a StepOnePlus thermocycler (Thermo Fisher Scientific, USA) with HiPi Real-Time PCR 2x Master Mix (SYBR green) according to the manufacturer’s instructions.

    Expressing:

    Article Title: L-Ferritin Binding to Scara5: A New Iron Traffic Pathway Potentially Implicated in Retinopathy
    Article Snippet: Paragraph title: Quantitative real-time PCR analysis of gene expression ... Gene relative quantification was carried out using MicroAmp Optical 96-well plates (Applied Biosystems) in a StepOnePlus thermocycler (Applied Biosystems) in standard cycling conditions.

    Article Title: Corticospinal-specific HCN expression in mouse motor cortex: Ih-dependent synaptic integration as a candidate microcircuit mechanism involved in motor control
    Article Snippet: Real-time PCR was performed using SYBR Green I on a StepOnePlus thermocycler (Applied Biosystems, Foster City, CA). .. A relative quantification method (ΔΔ comparative threshold method) ( ) was used to quantify expression levels of HCN1, HCN2, and Trip8b in cortical neurons with GAPDH as an endogenous reference gene.

    Article Title: Modeling appendicular skeletal cartilage development with modified high-density micromass cultures of adult human bone marrow-derived mesenchymal progenitor cells
    Article Snippet: Paragraph title: Gene expression analysis ... Quantitative real-time PCR was performed using a StepOnePlus thermocycler (Applied Biosystems) and SYBR Green Reaction Mix (Applied Biosystems).

    Article Title: Extracellular Vesicles Released by Leishmania (Leishmania) amazonensis Promote Disease Progression and Induce the Production of Different Cytokines in Macrophages and B-1 Cells
    Article Snippet: Paragraph title: Cytokine Expression ... The real-time PCR reactions (qRT-PCR) were performed using the SYBR Green Real-Time PCR Master Mix (Applied Biosystems, Thermo Fisher Scientific) and the StepOnePlus thermocycler (Applied Biosystems) with equal amounts of each cDNA.

    Article Title: Strain-Specific Regulation of Striatal Phenotype in Drd2-eGFP BAC Transgenic Mice
    Article Snippet: Paragraph title: Expression profiling ... Real-time PCR was performed using Fast SYBR Mastermix (Applied Biosystems) on a StepOnePlus thermocycler (Applied Biosystems).

    Article Title: In smokers, Sonic hedgehog modulates pulmonary endothelial function through vascular endothelial growth factor
    Article Snippet: .. After initial denaturation at 95 °C for 10 min, 20 ng of cDNA were amplified (using Gene Expression Master Mix, Life Technologies) in 40 annealing/extension cycles (95 °C for 15 s and 60 °C for 1 min) in a StepOnePlus thermocycler (Life Technologies). .. The sample’s fluorescence was measured after each cycle, and the threshold cycle (Ct) of the real-time PCR was defined as the point at which a fluorescence signal corresponding to the amplification of a PCR product was detectable.

    Transplantation Assay:

    Article Title: Fibroblast Growth Factor Receptor Signaling Is Essential for Normal Mammary Gland Development and Stem Cell Function
    Article Snippet: Cells for transplantation were never cultured on plastic to prevent loss of MaSCs and subsequent differentiation. .. RNA was collected through extraction with Trizol reagent (Invitrogen, Life Technologies, Carlsbad, CA) and cDNA templates were generated using a SuperScript III kit and 1 μ g RNA per sample according to the manufacturer’s protocol (Invitrogen, Carlsbad, CA). qPCR reactions were run using SYBR Green reagent (Applied Biosystems, Carlsbad, CA) on a StepOnePlus thermocycler (Applied Biosystems, Life Technologies, Carlsbad, CA) and fold changes were calculated using the Comparative CT (ΔΔCT) method and Step One software v2.0.1, Applied Biosystems, Life Technologies, Carlsbad, CA.

    Serial Dilution:

    Article Title: Control of mitotic chromosome condensation by the fission yeast transcription factor Zas1
    Article Snippet: Transcript abundance was measured by qPCR using SYBR Green Mix (Thermo Fisher) on a StepOnePlus thermocycler (Applied Biosystems) using primer pairs described in Table S2, and for act1 as described previously ( ). .. For each qPCR run, primer efficiency was determined by RT reaction serial dilution.

    SYBR Green Assay:

    Article Title: L-Ferritin Binding to Scara5: A New Iron Traffic Pathway Potentially Implicated in Retinopathy
    Article Snippet: Gene relative quantification was carried out using MicroAmp Optical 96-well plates (Applied Biosystems) in a StepOnePlus thermocycler (Applied Biosystems) in standard cycling conditions. .. The 12.5 µl PCR reaction mixtures contained 6.25 µl of 2×Power SYBR Green PCR Master Mix (Applied Biosystems), 160 nM of each primer, and 2 µl of diluted cDNA as template.

    Article Title: Corticospinal-specific HCN expression in mouse motor cortex: Ih-dependent synaptic integration as a candidate microcircuit mechanism involved in motor control
    Article Snippet: .. Real-time PCR was performed using SYBR Green I on a StepOnePlus thermocycler (Applied Biosystems, Foster City, CA). .. The thermal cycling conditions were comprised of an initial denaturing step at 95°C for 3 min and 40 cycles at 95°C for 15 s, 60°C for 60 s, and 72°C for 30 s. The PCR cycle threshold values were measured within the exponential phase of the PCR reaction using StepOnePlus software (version 2.1, Applied Biosystems).

    Article Title: Modeling appendicular skeletal cartilage development with modified high-density micromass cultures of adult human bone marrow-derived mesenchymal progenitor cells
    Article Snippet: .. Quantitative real-time PCR was performed using a StepOnePlus thermocycler (Applied Biosystems) and SYBR Green Reaction Mix (Applied Biosystems). .. Expression of aggrecan (ACAN ) and collagen type II (COL2A1 ) was quantified to analyze chondrogenic differentiation.

    Article Title: Deficient Vitamin E Uptake During Development Impairs Neural Tube Closure in Mice Lacking Lipoprotein Receptor SR-BI
    Article Snippet: .. The resulting cDNA was amplified by real time PCR with a StepOnePlus thermocycler (Applied Biosystems, CA) using the PowerUp SYBR Green master mix (Thermo, MA) and 100 nM of each primer. .. The primers, annealing temperatures and amplification efficiencies are listed in Supplementary Table .

    Article Title: Fibroblast Growth Factor Receptor Signaling Is Essential for Normal Mammary Gland Development and Stem Cell Function
    Article Snippet: .. RNA was collected through extraction with Trizol reagent (Invitrogen, Life Technologies, Carlsbad, CA) and cDNA templates were generated using a SuperScript III kit and 1 μ g RNA per sample according to the manufacturer’s protocol (Invitrogen, Carlsbad, CA). qPCR reactions were run using SYBR Green reagent (Applied Biosystems, Carlsbad, CA) on a StepOnePlus thermocycler (Applied Biosystems, Life Technologies, Carlsbad, CA) and fold changes were calculated using the Comparative CT (ΔΔCT) method and Step One software v2.0.1, Applied Biosystems, Life Technologies, Carlsbad, CA. ..

    Article Title: Control of mitotic chromosome condensation by the fission yeast transcription factor Zas1
    Article Snippet: .. Transcript abundance was measured by qPCR using SYBR Green Mix (Thermo Fisher) on a StepOnePlus thermocycler (Applied Biosystems) using primer pairs described in Table S2, and for act1 as described previously ( ). .. For each qPCR run, primer efficiency was determined by RT reaction serial dilution.

    Article Title: Extracellular Vesicles Released by Leishmania (Leishmania) amazonensis Promote Disease Progression and Induce the Production of Different Cytokines in Macrophages and B-1 Cells
    Article Snippet: .. The real-time PCR reactions (qRT-PCR) were performed using the SYBR Green Real-Time PCR Master Mix (Applied Biosystems, Thermo Fisher Scientific) and the StepOnePlus thermocycler (Applied Biosystems) with equal amounts of each cDNA. ..

    Article Title: Validation of reference genes for quantitative real-time PCR (qPCR) analysis of Actinobacillus suis
    Article Snippet: Three technical replicates were done per sample in a total reaction volume of 10 μL, which contained 5 μL of PerfeCTa® SYBR® Green FastMix® (Quanta BioSciences, Inc., Gaithersburg, MD), 2.5 μL of a forward/reverse primer mix with 1.6 μM of each primer, and 2.5 μL of cDNA diluted 1:15. .. Real-time PCR was done in a StepOnePlus Thermocycler (Applied Biosystems, Foster City, CA) using a program with an initial denaturation step at 95°C for 30 seconds followed by 40 cycles of 95°C for 3 seconds and 60°C for 30 seconds.

    Article Title: Molecular assessment of trematode co-infection and intraspecific competition in molluscan intermediate hosts
    Article Snippet: .. Real-time PCR and melt curve analyses were performed with Power SYBR Green MasterMix chemistry on a StepOnePlus thermocycler (Applied Biosystems). .. Following an initial 10-minute denaturation and Taq activation at 95°C, reactions were cycled as follows, using a “stepdown” scheme to encourage reaction stringency: 2 cycles of 15s at 95°C and 60s at 60°C, 2 cycles of 15s at 95°C and 60s at 59°C, and 40 cycles of 15s at 95°C and 60s at 58°C.

    Article Title: Biosynthesis of organic photosensitizer Zn-porphyrin by diphtheria toxin repressor (DtxR)-mediated global upregulation of engineered heme biosynthesis pathway in Corynebacterium glutamicum
    Article Snippet: .. All oligonucleotides for qRT-PCR were designed based on target gene sequence (Supplementary Table ). qRT-PCR analysis was determined using a StepOnePlus thermocycler (Thermo Fisher Scientific, USA) with HiPi Real-Time PCR 2x Master Mix (SYBR green) according to the manufacturer’s instructions. ..

    Article Title: Local TNFR1 signaling licenses murine neutrophils for increased TLR-dependent cytokine and eicosanoid production
    Article Snippet: .. Neutrophils were lysed in RNAzol (MRC) and RNA was purified according to manufacturer’s instructions and concentrated using on a column (RNA Clean and Concentrator, Zymo Research). cDNA was prepared with iScript cDNA synthesis kit (Biorad), and quantitative PCR was performed with SYBR Green Master mix (Biorad) on a StepOnePlus thermocycler (Applied Biosystems). .. Primers were obtained from PrimerBank ( ) and synthesized by IDT (see ).

    Article Title: Inhibition of HER2 Increases Jagged1-dependent Breast Cancer Stem Cells: Role for Membrane Jagged1
    Article Snippet: In a 96 well plate, 2.5 μL of prepared cDNA was added to 22.5 μL of a mastermix-containing 2X SYBR Green Universal Master Mix, and 50 μM forward and reverse primers for the intended target. .. A melt curve was added after completion of the 40 cycles set by the StepOnePlus thermocycler manufacturer (Applied Biosystems, Foster City, CA).

    Cell Culture:

    Article Title: Fibroblast Growth Factor Receptor Signaling Is Essential for Normal Mammary Gland Development and Stem Cell Function
    Article Snippet: Cells for transplantation were never cultured on plastic to prevent loss of MaSCs and subsequent differentiation. .. RNA was collected through extraction with Trizol reagent (Invitrogen, Life Technologies, Carlsbad, CA) and cDNA templates were generated using a SuperScript III kit and 1 μ g RNA per sample according to the manufacturer’s protocol (Invitrogen, Carlsbad, CA). qPCR reactions were run using SYBR Green reagent (Applied Biosystems, Carlsbad, CA) on a StepOnePlus thermocycler (Applied Biosystems, Life Technologies, Carlsbad, CA) and fold changes were calculated using the Comparative CT (ΔΔCT) method and Step One software v2.0.1, Applied Biosystems, Life Technologies, Carlsbad, CA.

    Article Title: Control of mitotic chromosome condensation by the fission yeast transcription factor Zas1
    Article Snippet: Cells of the desired genotype from three independent tetrads were cultured in YE5S at 25°C to OD600 0.8–1.0, and total RNA was isolated from 7.5 × 107 cells by phenol/chloroform extraction. .. Transcript abundance was measured by qPCR using SYBR Green Mix (Thermo Fisher) on a StepOnePlus thermocycler (Applied Biosystems) using primer pairs described in Table S2, and for act1 as described previously ( ).

    Generated:

    Article Title: Fibroblast Growth Factor Receptor Signaling Is Essential for Normal Mammary Gland Development and Stem Cell Function
    Article Snippet: .. RNA was collected through extraction with Trizol reagent (Invitrogen, Life Technologies, Carlsbad, CA) and cDNA templates were generated using a SuperScript III kit and 1 μ g RNA per sample according to the manufacturer’s protocol (Invitrogen, Carlsbad, CA). qPCR reactions were run using SYBR Green reagent (Applied Biosystems, Carlsbad, CA) on a StepOnePlus thermocycler (Applied Biosystems, Life Technologies, Carlsbad, CA) and fold changes were calculated using the Comparative CT (ΔΔCT) method and Step One software v2.0.1, Applied Biosystems, Life Technologies, Carlsbad, CA. ..

    Sequencing:

    Article Title: L-Ferritin Binding to Scara5: A New Iron Traffic Pathway Potentially Implicated in Retinopathy
    Article Snippet: Sequence homology searches were performed using the Basic Local Alignment Search Tool (Blast) “ http://www.ncbi.nlm.nih.gov/blast ”, to confirm gene identity of amplified fragments and that these primers matched only the sequence to which they were designed. .. Gene relative quantification was carried out using MicroAmp Optical 96-well plates (Applied Biosystems) in a StepOnePlus thermocycler (Applied Biosystems) in standard cycling conditions.

    Article Title: Biosynthesis of organic photosensitizer Zn-porphyrin by diphtheria toxin repressor (DtxR)-mediated global upregulation of engineered heme biosynthesis pathway in Corynebacterium glutamicum
    Article Snippet: .. All oligonucleotides for qRT-PCR were designed based on target gene sequence (Supplementary Table ). qRT-PCR analysis was determined using a StepOnePlus thermocycler (Thermo Fisher Scientific, USA) with HiPi Real-Time PCR 2x Master Mix (SYBR green) according to the manufacturer’s instructions. ..

    Article Title: Development of Novel Single-Chain Antibodies against the Hydrophobic HPV-16 E5 Protein
    Article Snippet: The primers for each sequence were 59-CTTGGGCACCGAAGAAACAC-39 (nucleotides 3438 to 3457) and 59-TTGGTCACGTTGCCATTCAC-39 (nucleotides 3770 to 3789) for the E2 gene and 59-AAGGGCGTAACCGAAATCGGT-39 (nucleotides 26 to 46) and 59-CATATACCTCACGTCGCAG-39 (nucleotides 215 to 233) for the E6 gene. .. The reaction was performed with Evagreen qPCR mastermix-Rox (ABM, Richmond, BC, Canada), following the manufacturer's instructions in a StepOnePlus thermocycler (Applied Biosystems).

    Fluorescence:

    Article Title: Molecular assessment of trematode co-infection and intraspecific competition in molluscan intermediate hosts
    Article Snippet: Real-time PCR and melt curve analyses were performed with Power SYBR Green MasterMix chemistry on a StepOnePlus thermocycler (Applied Biosystems). .. Fluorescence levels of the SYBR Green dye relative to ROX passive reference dye were assessed in real time at each 60-second annealing step during thermal cycling and then again with every 0.3°C increase during the melt curve analysis.

    Article Title: In smokers, Sonic hedgehog modulates pulmonary endothelial function through vascular endothelial growth factor
    Article Snippet: After initial denaturation at 95 °C for 10 min, 20 ng of cDNA were amplified (using Gene Expression Master Mix, Life Technologies) in 40 annealing/extension cycles (95 °C for 15 s and 60 °C for 1 min) in a StepOnePlus thermocycler (Life Technologies). .. The sample’s fluorescence was measured after each cycle, and the threshold cycle (Ct) of the real-time PCR was defined as the point at which a fluorescence signal corresponding to the amplification of a PCR product was detectable.

    Isolation:

    Article Title: Corticospinal-specific HCN expression in mouse motor cortex: Ih-dependent synaptic integration as a candidate microcircuit mechanism involved in motor control
    Article Snippet: Paragraph title: RNA Isolation and Real-Time PCR Quantification ... Real-time PCR was performed using SYBR Green I on a StepOnePlus thermocycler (Applied Biosystems, Foster City, CA).

    Article Title: Fibroblast Growth Factor Receptor Signaling Is Essential for Normal Mammary Gland Development and Stem Cell Function
    Article Snippet: Paragraph title: RNA Isolation and Quantitative Reverse Transcription-PCR ... RNA was collected through extraction with Trizol reagent (Invitrogen, Life Technologies, Carlsbad, CA) and cDNA templates were generated using a SuperScript III kit and 1 μ g RNA per sample according to the manufacturer’s protocol (Invitrogen, Carlsbad, CA). qPCR reactions were run using SYBR Green reagent (Applied Biosystems, Carlsbad, CA) on a StepOnePlus thermocycler (Applied Biosystems, Life Technologies, Carlsbad, CA) and fold changes were calculated using the Comparative CT (ΔΔCT) method and Step One software v2.0.1, Applied Biosystems, Life Technologies, Carlsbad, CA.

    Article Title: Control of mitotic chromosome condensation by the fission yeast transcription factor Zas1
    Article Snippet: Cells of the desired genotype from three independent tetrads were cultured in YE5S at 25°C to OD600 0.8–1.0, and total RNA was isolated from 7.5 × 107 cells by phenol/chloroform extraction. .. Transcript abundance was measured by qPCR using SYBR Green Mix (Thermo Fisher) on a StepOnePlus thermocycler (Applied Biosystems) using primer pairs described in Table S2, and for act1 as described previously ( ).

    Article Title: Strain-Specific Regulation of Striatal Phenotype in Drd2-eGFP BAC Transgenic Mice
    Article Snippet: Total RNA was isolated using RNeasy micro kit (Qiagen). cDNA was synthesized using qScript cDNA Supermix (Quanta Biosciences). .. Real-time PCR was performed using Fast SYBR Mastermix (Applied Biosystems) on a StepOnePlus thermocycler (Applied Biosystems).

    Article Title: In smokers, Sonic hedgehog modulates pulmonary endothelial function through vascular endothelial growth factor
    Article Snippet: Paragraph title: RNA isolation and reverse transcriptase – quantitative polymerase chain reaction (RT-qPCR) analysis ... After initial denaturation at 95 °C for 10 min, 20 ng of cDNA were amplified (using Gene Expression Master Mix, Life Technologies) in 40 annealing/extension cycles (95 °C for 15 s and 60 °C for 1 min) in a StepOnePlus thermocycler (Life Technologies).

    Negative Control:

    Article Title: Extracellular Vesicles Released by Leishmania (Leishmania) amazonensis Promote Disease Progression and Induce the Production of Different Cytokines in Macrophages and B-1 Cells
    Article Snippet: The real-time PCR reactions (qRT-PCR) were performed using the SYBR Green Real-Time PCR Master Mix (Applied Biosystems, Thermo Fisher Scientific) and the StepOnePlus thermocycler (Applied Biosystems) with equal amounts of each cDNA. .. The cycling parameters were 10 min at 50°C for enzyme activation, denaturation for 5 min at 95°C, and 40 cycles of 95°C for 30 s and 60°C for 1 min. A non-template control was included in each reaction as a negative control to enable the detection of contamination.

    Purification:

    Article Title: Modeling appendicular skeletal cartilage development with modified high-density micromass cultures of adult human bone marrow-derived mesenchymal progenitor cells
    Article Snippet: Gene expression analysis Total RNA was extracted using Trizol (Invitrogen) and purified with the RNeasy Plus mini kit (Qiagen). cDNA was reverse transcribed using the SuperScript IV kit (Invitrogen). .. Quantitative real-time PCR was performed using a StepOnePlus thermocycler (Applied Biosystems) and SYBR Green Reaction Mix (Applied Biosystems).

    Article Title: Deficient Vitamin E Uptake During Development Impairs Neural Tube Closure in Mice Lacking Lipoprotein Receptor SR-BI
    Article Snippet: Purified RNA (500 ng) was used for retrotranscription with the iScript RT Supermix (Biorad, CA). .. The resulting cDNA was amplified by real time PCR with a StepOnePlus thermocycler (Applied Biosystems, CA) using the PowerUp SYBR Green master mix (Thermo, MA) and 100 nM of each primer.

    Article Title: Local TNFR1 signaling licenses murine neutrophils for increased TLR-dependent cytokine and eicosanoid production
    Article Snippet: .. Neutrophils were lysed in RNAzol (MRC) and RNA was purified according to manufacturer’s instructions and concentrated using on a column (RNA Clean and Concentrator, Zymo Research). cDNA was prepared with iScript cDNA synthesis kit (Biorad), and quantitative PCR was performed with SYBR Green Master mix (Biorad) on a StepOnePlus thermocycler (Applied Biosystems). .. Primers were obtained from PrimerBank ( ) and synthesized by IDT (see ).

    Polymerase Chain Reaction:

    Article Title: L-Ferritin Binding to Scara5: A New Iron Traffic Pathway Potentially Implicated in Retinopathy
    Article Snippet: PCR products were sequenced and homology searches were performed with Blast. .. Gene relative quantification was carried out using MicroAmp Optical 96-well plates (Applied Biosystems) in a StepOnePlus thermocycler (Applied Biosystems) in standard cycling conditions.

    Article Title: Corticospinal-specific HCN expression in mouse motor cortex: Ih-dependent synaptic integration as a candidate microcircuit mechanism involved in motor control
    Article Snippet: Real-time PCR was performed using SYBR Green I on a StepOnePlus thermocycler (Applied Biosystems, Foster City, CA). .. The thermal cycling conditions were comprised of an initial denaturing step at 95°C for 3 min and 40 cycles at 95°C for 15 s, 60°C for 60 s, and 72°C for 30 s. The PCR cycle threshold values were measured within the exponential phase of the PCR reaction using StepOnePlus software (version 2.1, Applied Biosystems).

    Article Title: Control of mitotic chromosome condensation by the fission yeast transcription factor Zas1
    Article Snippet: Paragraph title: RT–quantitative PCR (qPCR) ... Transcript abundance was measured by qPCR using SYBR Green Mix (Thermo Fisher) on a StepOnePlus thermocycler (Applied Biosystems) using primer pairs described in Table S2, and for act1 as described previously ( ).

    Article Title: Strain-Specific Regulation of Striatal Phenotype in Drd2-eGFP BAC Transgenic Mice
    Article Snippet: Real-time PCR was performed using Fast SYBR Mastermix (Applied Biosystems) on a StepOnePlus thermocycler (Applied Biosystems). .. The thermal cycling conditions comprised of an initial denaturing step at 95°C for 20 s, and 40 cycles at 95°C for 3 s, 60°C for 30 s. The PCR cycle threshold (Ct) values were measured within the exponential phase of the PCR reaction using StepOnePlus software version 2.1 (Applied Biosystems).

    Article Title: In smokers, Sonic hedgehog modulates pulmonary endothelial function through vascular endothelial growth factor
    Article Snippet: After initial denaturation at 95 °C for 10 min, 20 ng of cDNA were amplified (using Gene Expression Master Mix, Life Technologies) in 40 annealing/extension cycles (95 °C for 15 s and 60 °C for 1 min) in a StepOnePlus thermocycler (Life Technologies). .. The sample’s fluorescence was measured after each cycle, and the threshold cycle (Ct) of the real-time PCR was defined as the point at which a fluorescence signal corresponding to the amplification of a PCR product was detectable.

    Article Title: Inhibition of HER2 Increases Jagged1-dependent Breast Cancer Stem Cells: Role for Membrane Jagged1
    Article Snippet: The plate was then sealed to prevent evaporation and quantitative Real Time PCR was performed using following parameters: the initial denaturation was for 10 minutes at 95°C; PCR cycling for 40 cycles was carried out for 10 seconds at 95°C and 45 seconds at 60°C. .. A melt curve was added after completion of the 40 cycles set by the StepOnePlus thermocycler manufacturer (Applied Biosystems, Foster City, CA).

    Article Title: Development of Novel Single-Chain Antibodies against the Hydrophobic HPV-16 E5 Protein
    Article Snippet: For detection of integration, a multiplex PCR for the HPV-16 E2 and HPV-16 E6 genes was performed. .. The reaction was performed with Evagreen qPCR mastermix-Rox (ABM, Richmond, BC, Canada), following the manufacturer's instructions in a StepOnePlus thermocycler (Applied Biosystems).

    Software:

    Article Title: Corticospinal-specific HCN expression in mouse motor cortex: Ih-dependent synaptic integration as a candidate microcircuit mechanism involved in motor control
    Article Snippet: Real-time PCR was performed using SYBR Green I on a StepOnePlus thermocycler (Applied Biosystems, Foster City, CA). .. The thermal cycling conditions were comprised of an initial denaturing step at 95°C for 3 min and 40 cycles at 95°C for 15 s, 60°C for 60 s, and 72°C for 30 s. The PCR cycle threshold values were measured within the exponential phase of the PCR reaction using StepOnePlus software (version 2.1, Applied Biosystems).

    Article Title: Fibroblast Growth Factor Receptor Signaling Is Essential for Normal Mammary Gland Development and Stem Cell Function
    Article Snippet: .. RNA was collected through extraction with Trizol reagent (Invitrogen, Life Technologies, Carlsbad, CA) and cDNA templates were generated using a SuperScript III kit and 1 μ g RNA per sample according to the manufacturer’s protocol (Invitrogen, Carlsbad, CA). qPCR reactions were run using SYBR Green reagent (Applied Biosystems, Carlsbad, CA) on a StepOnePlus thermocycler (Applied Biosystems, Life Technologies, Carlsbad, CA) and fold changes were calculated using the Comparative CT (ΔΔCT) method and Step One software v2.0.1, Applied Biosystems, Life Technologies, Carlsbad, CA. ..

    Article Title: Control of mitotic chromosome condensation by the fission yeast transcription factor Zas1
    Article Snippet: Transcript abundance was measured by qPCR using SYBR Green Mix (Thermo Fisher) on a StepOnePlus thermocycler (Applied Biosystems) using primer pairs described in Table S2, and for act1 as described previously ( ). .. CT values were calculated with StepOne software (Applied Biosystems).

    Article Title: Strain-Specific Regulation of Striatal Phenotype in Drd2-eGFP BAC Transgenic Mice
    Article Snippet: Real-time PCR was performed using Fast SYBR Mastermix (Applied Biosystems) on a StepOnePlus thermocycler (Applied Biosystems). .. The thermal cycling conditions comprised of an initial denaturing step at 95°C for 20 s, and 40 cycles at 95°C for 3 s, 60°C for 30 s. The PCR cycle threshold (Ct) values were measured within the exponential phase of the PCR reaction using StepOnePlus software version 2.1 (Applied Biosystems).

    Electrophoresis:

    Article Title: Extracellular Vesicles Released by Leishmania (Leishmania) amazonensis Promote Disease Progression and Induce the Production of Different Cytokines in Macrophages and B-1 Cells
    Article Snippet: Only RNA samples with reading ranges of 1.8–2.0 at 260/280 nm and 260/230 nm were subjected to electrophoresis in 1.5% agarose gels to assess RNA integrity. .. The real-time PCR reactions (qRT-PCR) were performed using the SYBR Green Real-Time PCR Master Mix (Applied Biosystems, Thermo Fisher Scientific) and the StepOnePlus thermocycler (Applied Biosystems) with equal amounts of each cDNA.

    Article Title: In smokers, Sonic hedgehog modulates pulmonary endothelial function through vascular endothelial growth factor
    Article Snippet: The amount of RNA extracted was estimated by spectrophotometry at 260 nm (Biowave DNA; Biochrom, Cambridge, UK) and the quality of the preparation was assessed in a microfluidic electrophoresis system (RNA Standard Sensitivity kits for Experion, BioRad, Marnes-la-Coquette, France). .. After initial denaturation at 95 °C for 10 min, 20 ng of cDNA were amplified (using Gene Expression Master Mix, Life Technologies) in 40 annealing/extension cycles (95 °C for 15 s and 60 °C for 1 min) in a StepOnePlus thermocycler (Life Technologies).

    Multiplex Assay:

    Article Title: Development of Novel Single-Chain Antibodies against the Hydrophobic HPV-16 E5 Protein
    Article Snippet: For detection of integration, a multiplex PCR for the HPV-16 E2 and HPV-16 E6 genes was performed. .. The reaction was performed with Evagreen qPCR mastermix-Rox (ABM, Richmond, BC, Canada), following the manufacturer's instructions in a StepOnePlus thermocycler (Applied Biosystems).

    Agarose Gel Electrophoresis:

    Article Title: L-Ferritin Binding to Scara5: A New Iron Traffic Pathway Potentially Implicated in Retinopathy
    Article Snippet: Gene relative quantification was carried out using MicroAmp Optical 96-well plates (Applied Biosystems) in a StepOnePlus thermocycler (Applied Biosystems) in standard cycling conditions. .. The primer specificity and the formation of primer-dimers were confirmed by melt curve analysis and agarose gel electrophoresis.

    Spectrophotometry:

    Article Title: Extracellular Vesicles Released by Leishmania (Leishmania) amazonensis Promote Disease Progression and Induce the Production of Different Cytokines in Macrophages and B-1 Cells
    Article Snippet: The total RNA was quantified by UV absorption using a spectrophotometer (Nanodrop 2000c, Thermo Fisher). .. The real-time PCR reactions (qRT-PCR) were performed using the SYBR Green Real-Time PCR Master Mix (Applied Biosystems, Thermo Fisher Scientific) and the StepOnePlus thermocycler (Applied Biosystems) with equal amounts of each cDNA.

    Article Title: In smokers, Sonic hedgehog modulates pulmonary endothelial function through vascular endothelial growth factor
    Article Snippet: The amount of RNA extracted was estimated by spectrophotometry at 260 nm (Biowave DNA; Biochrom, Cambridge, UK) and the quality of the preparation was assessed in a microfluidic electrophoresis system (RNA Standard Sensitivity kits for Experion, BioRad, Marnes-la-Coquette, France). .. After initial denaturation at 95 °C for 10 min, 20 ng of cDNA were amplified (using Gene Expression Master Mix, Life Technologies) in 40 annealing/extension cycles (95 °C for 15 s and 60 °C for 1 min) in a StepOnePlus thermocycler (Life Technologies).

    Evaporation:

    Article Title: Inhibition of HER2 Increases Jagged1-dependent Breast Cancer Stem Cells: Role for Membrane Jagged1
    Article Snippet: The plate was then sealed to prevent evaporation and quantitative Real Time PCR was performed using following parameters: the initial denaturation was for 10 minutes at 95°C; PCR cycling for 40 cycles was carried out for 10 seconds at 95°C and 45 seconds at 60°C. .. A melt curve was added after completion of the 40 cycles set by the StepOnePlus thermocycler manufacturer (Applied Biosystems, Foster City, CA).

    Activation Assay:

    Article Title: Extracellular Vesicles Released by Leishmania (Leishmania) amazonensis Promote Disease Progression and Induce the Production of Different Cytokines in Macrophages and B-1 Cells
    Article Snippet: The real-time PCR reactions (qRT-PCR) were performed using the SYBR Green Real-Time PCR Master Mix (Applied Biosystems, Thermo Fisher Scientific) and the StepOnePlus thermocycler (Applied Biosystems) with equal amounts of each cDNA. .. The cycling parameters were 10 min at 50°C for enzyme activation, denaturation for 5 min at 95°C, and 40 cycles of 95°C for 30 s and 60°C for 1 min. A non-template control was included in each reaction as a negative control to enable the detection of contamination.

    Article Title: Molecular assessment of trematode co-infection and intraspecific competition in molluscan intermediate hosts
    Article Snippet: Real-time PCR and melt curve analyses were performed with Power SYBR Green MasterMix chemistry on a StepOnePlus thermocycler (Applied Biosystems). .. Following an initial 10-minute denaturation and Taq activation at 95°C, reactions were cycled as follows, using a “stepdown” scheme to encourage reaction stringency: 2 cycles of 15s at 95°C and 60s at 60°C, 2 cycles of 15s at 95°C and 60s at 59°C, and 40 cycles of 15s at 95°C and 60s at 58°C.

    FACS:

    Article Title: Corticospinal-specific HCN expression in mouse motor cortex: Ih-dependent synaptic integration as a candidate microcircuit mechanism involved in motor control
    Article Snippet: Total RNA was isolated from FACS-isolated cells using TRIzol (Invitrogen). .. Real-time PCR was performed using SYBR Green I on a StepOnePlus thermocycler (Applied Biosystems, Foster City, CA).

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