steponeplus real time pcr  (Thermo Fisher)


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    Structured Review

    Thermo Fisher steponeplus real time pcr
    Steponeplus Real Time Pcr, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 97/100, based on 57 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/steponeplus real time pcr/product/Thermo Fisher
    Average 97 stars, based on 57 article reviews
    Price from $9.99 to $1999.99
    steponeplus real time pcr - by Bioz Stars, 2020-04
    97/100 stars

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    Related Articles

    Countercurrent Chromatography:

    Article Title: Erastin Inhibits Septic Shock and Inflammatory Gene Expression via Suppression of the NF-κB Pathway
    Article Snippet: Triplicate reactions were performed using the StepOnePlus Real-Time PCR (Thermo). .. Primer sequences were as follows: β-actin (forward: 5′- TTG CTG ACA GGA TGC AGA AG-3′, reverse: 5′-ACA TCT GCT GGA AGG TGG AC-3′); iNOS (forward: 5′-CAC CTT GGA GTT CAC CCA GT-3′, reverse: 5′-TGG TCA CCT CCA ACA CAA GA-3′); COX2 (forward: 5′- GGC CAT GGA GTG GAC TTA AA-3′, reverse: 5′-ACC TCT CCA CCA ATG ACC TG-3′); IL-1β (forward: 5′-GAG CCC ATC CTC TGT GAC TC-3′, reverse: 5′-AGC TCA TAT GGG TCC GAC AG-3′); TNF-α (forward: 5′-ACG GCA TGG ATC TCA AAG AC-3′, reverse: 5′-AGA TAG CAA ATC GGC TGA CG-3′).

    Real-time Polymerase Chain Reaction:

    Article Title: Data on the role of miR-144 in regulating fetal hemoglobin production in retinal pigmented epithelial cells
    Article Snippet: .. The following two-step thermal cycling profile was used (StepOnePlus Real-Time PCR, Life Technologies, Grand Island, NY): Step I (cycling): 95 °C for 15 min, 94 °C for 15 s, 55 °C for 30 s and 70 °C for 30s for 40 cycles. .. Step II (melting curve): 60 °C for 15 s, 60 °C 1 min and 95 °C for 30 s. The template amplification was confirmed by melting curve analysis. miRNA expression of genes were normalized to 5S expression and fold change in expression was calculated by 2–ΔΔCt method.

    Article Title: Cross-Packaging and Capsid Mosaic Formation in Multiplexed AAV Libraries
    Article Snippet: .. Digested samples were then supplemented with 20 μL of 20 mg/mL proteinase K (740506; MACHEREY-NAGEL) and incubated at 70°C for 20 min. vgs were further extracted using the NucleoSpin RNA Virus kit (MACHEREY-NAGEL). qPCR was performed with a StepOnePlus Real-Time PCR (Life Technologies, Carlsbad, CA, USA). qPCR reactions were run in a final volume of 20 μL, containing the primers/probe PCR Master Mix (TaKaRa, Kusatsu, Japan) and 5 μL of template DNA. .. For ITR titration, the standard plasmid was prepared in accordance with D’Costa et al.

    Article Title: Data on the role of miR-144 in regulating fetal hemoglobin production in retinal pigmented epithelial cells
    Article Snippet: .. The following two-step thermal cycling profile was used (StepOnePlus Real-Time PCR, Life Technologies, Grand Island, NY): Step I (cycling): 95 °C for 5 min, 95 °C for 15 s, 60 °C for 30 s and 72 °C for 15s for 40 cycles. .. Step II (melting curve): 60 °C for 15 s, 60 °C 1 min and 95 °C for 30 s. The template amplification was confirmed by melting curve analysis. mRNA expression of genes were normalized to 18s expression and fold change in expression was calculated by 2–ΔΔCt method.

    Article Title: Erastin Inhibits Septic Shock and Inflammatory Gene Expression via Suppression of the NF-κB Pathway
    Article Snippet: .. Triplicate reactions were performed using the StepOnePlus Real-Time PCR (Thermo). .. Primer sequences were as follows: β-actin (forward: 5′- TTG CTG ACA GGA TGC AGA AG-3′, reverse: 5′-ACA TCT GCT GGA AGG TGG AC-3′); iNOS (forward: 5′-CAC CTT GGA GTT CAC CCA GT-3′, reverse: 5′-TGG TCA CCT CCA ACA CAA GA-3′); COX2 (forward: 5′- GGC CAT GGA GTG GAC TTA AA-3′, reverse: 5′-ACC TCT CCA CCA ATG ACC TG-3′); IL-1β (forward: 5′-GAG CCC ATC CTC TGT GAC TC-3′, reverse: 5′-AGC TCA TAT GGG TCC GAC AG-3′); TNF-α (forward: 5′-ACG GCA TGG ATC TCA AAG AC-3′, reverse: 5′-AGA TAG CAA ATC GGC TGA CG-3′).

    Article Title: Expression of Neurotrophic Factors, Tight Junction Proteins, and Cytokines According to the Irritable Bowel Syndrome Subtype and Sex
    Article Snippet: .. Real-time quantitative polymerase chain reaction was performed in triplicates using a StepOnePlus Real-time PCR (Applied Biosystems) with SYBR Premix Ex TaqTM (Takara Bio, Shiga, Japan) according to manufacturers’ instructions and protocols. ..

    Article Title: Hepatitis C Virus Enhances the Invasiveness of Hepatocellular Carcinoma via EGFR-Mediated Invadopodia Formation and Activation
    Article Snippet: .. Real-time PCR was carried out in a StepOnePlus Real-Time PCR (Applied Biosystems) using the SYBR-Green PCR Master Mix (Applied Biosystems) according to the manufacturer’s protocol with 25 ng of total genomic DNA. .. The percent of human cells in the mice tissues was calculated.

    Article Title: SOX9 expression decreases survival of patients with intrahepatic cholangiocarcinoma by conferring chemoresistance
    Article Snippet: .. For PCR amplification, 10.4 µL mixtures contained 5 µL (50 ng) template cDNA, 5 µL SYBR Green (4367659, Life Technologies) and 4 µM forward and reverse primer PCRs were run in triplicate and performed on a StepOnePlus Real-time PCR (Applied Biosystems). .. PCR amplification cycling conditions comprised 10 min polymerase activation at 95 °C and 40 cycles at 95 °C for 15 s and 60 °C for 1 min. A melting-curve analysis was performed for each PCR analysis.

    Amplification:

    Article Title: Data on the role of miR-144 in regulating fetal hemoglobin production in retinal pigmented epithelial cells
    Article Snippet: Step II (melting curve): 60 °C for 15 s, 60 °C 1 min and 95 °C for 30 s. The template amplification was confirmed by melting curve analysis. mRNA expression of genes were normalized to 18s expression and fold change in expression was calculated by 2–ΔΔCt method. .. The following two-step thermal cycling profile was used (StepOnePlus Real-Time PCR, Life Technologies, Grand Island, NY): Step I (cycling): 95 °C for 15 min, 94 °C for 15 s, 55 °C for 30 s and 70 °C for 30s for 40 cycles.

    Article Title: Hepatitis C Virus Enhances the Invasiveness of Hepatocellular Carcinoma via EGFR-Mediated Invadopodia Formation and Activation
    Article Snippet: Mouse (Mo)-specific primers (forward: 5′-ttggttgagaagcagaaaca-3′, reverse: 5′-cacacagtcaagttcccaaa-3′) amplified a 181-bp fragment of a region located in 2F1–F3. .. Real-time PCR was carried out in a StepOnePlus Real-Time PCR (Applied Biosystems) using the SYBR-Green PCR Master Mix (Applied Biosystems) according to the manufacturer’s protocol with 25 ng of total genomic DNA.

    Article Title: SOX9 expression decreases survival of patients with intrahepatic cholangiocarcinoma by conferring chemoresistance
    Article Snippet: .. For PCR amplification, 10.4 µL mixtures contained 5 µL (50 ng) template cDNA, 5 µL SYBR Green (4367659, Life Technologies) and 4 µM forward and reverse primer PCRs were run in triplicate and performed on a StepOnePlus Real-time PCR (Applied Biosystems). .. PCR amplification cycling conditions comprised 10 min polymerase activation at 95 °C and 40 cycles at 95 °C for 15 s and 60 °C for 1 min. A melting-curve analysis was performed for each PCR analysis.

    Concentration Assay:

    Article Title: Tumor-suppressing potential of stingless bee propolis in in vitro and in vivo models of differentiated-type gastric adenocarcinoma
    Article Snippet: Quantitative real-time PCR (qRT-PCR) Gene expression profiles of human gastric cancer cell lines as well as the harvested mice stomach tissues were achieved using StepOnePlus Real-Time PCR (Applied Biosystems, CA, USA). .. For the former, cells at suitable density were seeded into a 6-well/flat-bottom microplate containing 2 ml culture medium and incubated for 48 hours with culture medium alone (control) or EEP with concentration corresponding to the calculated IC50 value (at 48 hours) for each respective gastric cancer cell lines.

    Isolation:

    Article Title: Data on the role of miR-144 in regulating fetal hemoglobin production in retinal pigmented epithelial cells
    Article Snippet: 2.4 Reverse transcription–quantitative polymerase chain reaction Total miRNA and RNA was isolated from RPE/eyecup of mice or ARPE-19 cells using miRNAeasy mini and RNAeasy kits respectively (Qiagen, USA). cDNA was prepared from total miRNA and RNA using the miScript RT (Qiagen, USA) and iScript cDNA Synthesis Kit (Bio-Rad) respectively and subjected to qPCR assays. .. The following two-step thermal cycling profile was used (StepOnePlus Real-Time PCR, Life Technologies, Grand Island, NY): Step I (cycling): 95 °C for 5 min, 95 °C for 15 s, 60 °C for 30 s and 72 °C for 15s for 40 cycles.

    Article Title: SOX9 expression decreases survival of patients with intrahepatic cholangiocarcinoma by conferring chemoresistance
    Article Snippet: Paragraph title: RNA isolation and quantitative real-time reverse transcription polymerase chain reaction ... For PCR amplification, 10.4 µL mixtures contained 5 µL (50 ng) template cDNA, 5 µL SYBR Green (4367659, Life Technologies) and 4 µM forward and reverse primer PCRs were run in triplicate and performed on a StepOnePlus Real-time PCR (Applied Biosystems).

    Article Title: Tumor-suppressing potential of stingless bee propolis in in vitro and in vivo models of differentiated-type gastric adenocarcinoma
    Article Snippet: Quantitative real-time PCR (qRT-PCR) Gene expression profiles of human gastric cancer cell lines as well as the harvested mice stomach tissues were achieved using StepOnePlus Real-Time PCR (Applied Biosystems, CA, USA). .. RNA was extracted in all samples using Nucleospin RNA isolation kit (Macherel-Nagel, Düren, Germany) according to the manufacturer’s instructions.

    Mouse Assay:

    Article Title: Data on the role of miR-144 in regulating fetal hemoglobin production in retinal pigmented epithelial cells
    Article Snippet: 2.4 Reverse transcription–quantitative polymerase chain reaction Total miRNA and RNA was isolated from RPE/eyecup of mice or ARPE-19 cells using miRNAeasy mini and RNAeasy kits respectively (Qiagen, USA). cDNA was prepared from total miRNA and RNA using the miScript RT (Qiagen, USA) and iScript cDNA Synthesis Kit (Bio-Rad) respectively and subjected to qPCR assays. .. The following two-step thermal cycling profile was used (StepOnePlus Real-Time PCR, Life Technologies, Grand Island, NY): Step I (cycling): 95 °C for 5 min, 95 °C for 15 s, 60 °C for 30 s and 72 °C for 15s for 40 cycles.

    Article Title: Hepatitis C Virus Enhances the Invasiveness of Hepatocellular Carcinoma via EGFR-Mediated Invadopodia Formation and Activation
    Article Snippet: Real-time PCR was carried out in a StepOnePlus Real-Time PCR (Applied Biosystems) using the SYBR-Green PCR Master Mix (Applied Biosystems) according to the manufacturer’s protocol with 25 ng of total genomic DNA. .. The percent of human cells in the mice tissues was calculated.

    Article Title: Tumor-suppressing potential of stingless bee propolis in in vitro and in vivo models of differentiated-type gastric adenocarcinoma
    Article Snippet: .. Quantitative real-time PCR (qRT-PCR) Gene expression profiles of human gastric cancer cell lines as well as the harvested mice stomach tissues were achieved using StepOnePlus Real-Time PCR (Applied Biosystems, CA, USA). .. For the former, cells at suitable density were seeded into a 6-well/flat-bottom microplate containing 2 ml culture medium and incubated for 48 hours with culture medium alone (control) or EEP with concentration corresponding to the calculated IC50 value (at 48 hours) for each respective gastric cancer cell lines.

    Quantitative RT-PCR:

    Article Title: Tumor-suppressing potential of stingless bee propolis in in vitro and in vivo models of differentiated-type gastric adenocarcinoma
    Article Snippet: .. Quantitative real-time PCR (qRT-PCR) Gene expression profiles of human gastric cancer cell lines as well as the harvested mice stomach tissues were achieved using StepOnePlus Real-Time PCR (Applied Biosystems, CA, USA). .. For the former, cells at suitable density were seeded into a 6-well/flat-bottom microplate containing 2 ml culture medium and incubated for 48 hours with culture medium alone (control) or EEP with concentration corresponding to the calculated IC50 value (at 48 hours) for each respective gastric cancer cell lines.

    Purification:

    Article Title: Expression of Neurotrophic Factors, Tight Junction Proteins, and Cytokines According to the Irritable Bowel Syndrome Subtype and Sex
    Article Snippet: Extracted RNA was purified using RNeasy mini kits (Qiagen, Valencia, CA, USA). .. Real-time quantitative polymerase chain reaction was performed in triplicates using a StepOnePlus Real-time PCR (Applied Biosystems) with SYBR Premix Ex TaqTM (Takara Bio, Shiga, Japan) according to manufacturers’ instructions and protocols.

    SYBR Green Assay:

    Article Title: Data on the role of miR-144 in regulating fetal hemoglobin production in retinal pigmented epithelial cells
    Article Snippet: The reaction volume of 20 μl for miRNA analysis contained 10.0 μl Quantitact SYBR green master mix (2X), 2 μl cDNA, 2 μl of universal primer, 2 μl of miRNA specific primer and 4 μl nuclease-free water. .. The following two-step thermal cycling profile was used (StepOnePlus Real-Time PCR, Life Technologies, Grand Island, NY): Step I (cycling): 95 °C for 15 min, 94 °C for 15 s, 55 °C for 30 s and 70 °C for 30s for 40 cycles.

    Article Title: Data on the role of miR-144 in regulating fetal hemoglobin production in retinal pigmented epithelial cells
    Article Snippet: The reaction volume of 20 μl for mRNA analysis contained 10.0 μl SYBR green master mix (2X), 1 μl cDNA, 1 μl of each primer and 7 μl nuclease-free water. .. The following two-step thermal cycling profile was used (StepOnePlus Real-Time PCR, Life Technologies, Grand Island, NY): Step I (cycling): 95 °C for 5 min, 95 °C for 15 s, 60 °C for 30 s and 72 °C for 15s for 40 cycles.

    Article Title: Hepatitis C Virus Enhances the Invasiveness of Hepatocellular Carcinoma via EGFR-Mediated Invadopodia Formation and Activation
    Article Snippet: .. Real-time PCR was carried out in a StepOnePlus Real-Time PCR (Applied Biosystems) using the SYBR-Green PCR Master Mix (Applied Biosystems) according to the manufacturer’s protocol with 25 ng of total genomic DNA. .. The percent of human cells in the mice tissues was calculated.

    Article Title: SOX9 expression decreases survival of patients with intrahepatic cholangiocarcinoma by conferring chemoresistance
    Article Snippet: .. For PCR amplification, 10.4 µL mixtures contained 5 µL (50 ng) template cDNA, 5 µL SYBR Green (4367659, Life Technologies) and 4 µM forward and reverse primer PCRs were run in triplicate and performed on a StepOnePlus Real-time PCR (Applied Biosystems). .. PCR amplification cycling conditions comprised 10 min polymerase activation at 95 °C and 40 cycles at 95 °C for 15 s and 60 °C for 1 min. A melting-curve analysis was performed for each PCR analysis.

    Activation Assay:

    Article Title: SOX9 expression decreases survival of patients with intrahepatic cholangiocarcinoma by conferring chemoresistance
    Article Snippet: For PCR amplification, 10.4 µL mixtures contained 5 µL (50 ng) template cDNA, 5 µL SYBR Green (4367659, Life Technologies) and 4 µM forward and reverse primer PCRs were run in triplicate and performed on a StepOnePlus Real-time PCR (Applied Biosystems). .. PCR amplification cycling conditions comprised 10 min polymerase activation at 95 °C and 40 cycles at 95 °C for 15 s and 60 °C for 1 min. A melting-curve analysis was performed for each PCR analysis.

    Incubation:

    Article Title: Cross-Packaging and Capsid Mosaic Formation in Multiplexed AAV Libraries
    Article Snippet: .. Digested samples were then supplemented with 20 μL of 20 mg/mL proteinase K (740506; MACHEREY-NAGEL) and incubated at 70°C for 20 min. vgs were further extracted using the NucleoSpin RNA Virus kit (MACHEREY-NAGEL). qPCR was performed with a StepOnePlus Real-Time PCR (Life Technologies, Carlsbad, CA, USA). qPCR reactions were run in a final volume of 20 μL, containing the primers/probe PCR Master Mix (TaKaRa, Kusatsu, Japan) and 5 μL of template DNA. .. For ITR titration, the standard plasmid was prepared in accordance with D’Costa et al.

    Article Title: Tumor-suppressing potential of stingless bee propolis in in vitro and in vivo models of differentiated-type gastric adenocarcinoma
    Article Snippet: Quantitative real-time PCR (qRT-PCR) Gene expression profiles of human gastric cancer cell lines as well as the harvested mice stomach tissues were achieved using StepOnePlus Real-Time PCR (Applied Biosystems, CA, USA). .. For the former, cells at suitable density were seeded into a 6-well/flat-bottom microplate containing 2 ml culture medium and incubated for 48 hours with culture medium alone (control) or EEP with concentration corresponding to the calculated IC50 value (at 48 hours) for each respective gastric cancer cell lines.

    Polymerase Chain Reaction:

    Article Title: Data on the role of miR-144 in regulating fetal hemoglobin production in retinal pigmented epithelial cells
    Article Snippet: Paragraph title: Reverse transcription–quantitative polymerase chain reaction ... The following two-step thermal cycling profile was used (StepOnePlus Real-Time PCR, Life Technologies, Grand Island, NY): Step I (cycling): 95 °C for 15 min, 94 °C for 15 s, 55 °C for 30 s and 70 °C for 30s for 40 cycles.

    Article Title: Cross-Packaging and Capsid Mosaic Formation in Multiplexed AAV Libraries
    Article Snippet: .. Digested samples were then supplemented with 20 μL of 20 mg/mL proteinase K (740506; MACHEREY-NAGEL) and incubated at 70°C for 20 min. vgs were further extracted using the NucleoSpin RNA Virus kit (MACHEREY-NAGEL). qPCR was performed with a StepOnePlus Real-Time PCR (Life Technologies, Carlsbad, CA, USA). qPCR reactions were run in a final volume of 20 μL, containing the primers/probe PCR Master Mix (TaKaRa, Kusatsu, Japan) and 5 μL of template DNA. .. For ITR titration, the standard plasmid was prepared in accordance with D’Costa et al.

    Article Title: Hepatitis C Virus Enhances the Invasiveness of Hepatocellular Carcinoma via EGFR-Mediated Invadopodia Formation and Activation
    Article Snippet: .. Real-time PCR was carried out in a StepOnePlus Real-Time PCR (Applied Biosystems) using the SYBR-Green PCR Master Mix (Applied Biosystems) according to the manufacturer’s protocol with 25 ng of total genomic DNA. .. The percent of human cells in the mice tissues was calculated.

    Article Title: SOX9 expression decreases survival of patients with intrahepatic cholangiocarcinoma by conferring chemoresistance
    Article Snippet: .. For PCR amplification, 10.4 µL mixtures contained 5 µL (50 ng) template cDNA, 5 µL SYBR Green (4367659, Life Technologies) and 4 µM forward and reverse primer PCRs were run in triplicate and performed on a StepOnePlus Real-time PCR (Applied Biosystems). .. PCR amplification cycling conditions comprised 10 min polymerase activation at 95 °C and 40 cycles at 95 °C for 15 s and 60 °C for 1 min. A melting-curve analysis was performed for each PCR analysis.

    Expressing:

    Article Title: Data on the role of miR-144 in regulating fetal hemoglobin production in retinal pigmented epithelial cells
    Article Snippet: Step II (melting curve): 60 °C for 15 s, 60 °C 1 min and 95 °C for 30 s. The template amplification was confirmed by melting curve analysis. mRNA expression of genes were normalized to 18s expression and fold change in expression was calculated by 2–ΔΔCt method. .. The following two-step thermal cycling profile was used (StepOnePlus Real-Time PCR, Life Technologies, Grand Island, NY): Step I (cycling): 95 °C for 15 min, 94 °C for 15 s, 55 °C for 30 s and 70 °C for 30s for 40 cycles.

    Article Title: Erastin Inhibits Septic Shock and Inflammatory Gene Expression via Suppression of the NF-κB Pathway
    Article Snippet: Triplicate reactions were performed using the StepOnePlus Real-Time PCR (Thermo). .. RNA quantity was normalized to β-actin content, and gene expression was quantified according to the 2−ΔCt method.

    Article Title: Expression of Neurotrophic Factors, Tight Junction Proteins, and Cytokines According to the Irritable Bowel Syndrome Subtype and Sex
    Article Snippet: Real-time quantitative polymerase chain reaction was performed in triplicates using a StepOnePlus Real-time PCR (Applied Biosystems) with SYBR Premix Ex TaqTM (Takara Bio, Shiga, Japan) according to manufacturers’ instructions and protocols. .. Expression levels of mRNAs for target genes were compared with endogenous control β-actin using the 2−ΔΔCt method.

    Article Title: Hepatitis C Virus Enhances the Invasiveness of Hepatocellular Carcinoma via EGFR-Mediated Invadopodia Formation and Activation
    Article Snippet: Real-time PCR was carried out in a StepOnePlus Real-Time PCR (Applied Biosystems) using the SYBR-Green PCR Master Mix (Applied Biosystems) according to the manufacturer’s protocol with 25 ng of total genomic DNA. .. To quantify the expression of specific genes in human cells that invaded into the mouse liver, total RNA was extracted from the left and right lobes of mouse livers using the RNeasy Mini kit and RNase-Free DNase Set.

    Article Title: Tumor-suppressing potential of stingless bee propolis in in vitro and in vivo models of differentiated-type gastric adenocarcinoma
    Article Snippet: .. Quantitative real-time PCR (qRT-PCR) Gene expression profiles of human gastric cancer cell lines as well as the harvested mice stomach tissues were achieved using StepOnePlus Real-Time PCR (Applied Biosystems, CA, USA). .. For the former, cells at suitable density were seeded into a 6-well/flat-bottom microplate containing 2 ml culture medium and incubated for 48 hours with culture medium alone (control) or EEP with concentration corresponding to the calculated IC50 value (at 48 hours) for each respective gastric cancer cell lines.

    CTG Assay:

    Article Title: Erastin Inhibits Septic Shock and Inflammatory Gene Expression via Suppression of the NF-κB Pathway
    Article Snippet: Triplicate reactions were performed using the StepOnePlus Real-Time PCR (Thermo). .. Primer sequences were as follows: β-actin (forward: 5′- TTG CTG ACA GGA TGC AGA AG-3′, reverse: 5′-ACA TCT GCT GGA AGG TGG AC-3′); iNOS (forward: 5′-CAC CTT GGA GTT CAC CCA GT-3′, reverse: 5′-TGG TCA CCT CCA ACA CAA GA-3′); COX2 (forward: 5′- GGC CAT GGA GTG GAC TTA AA-3′, reverse: 5′-ACC TCT CCA CCA ATG ACC TG-3′); IL-1β (forward: 5′-GAG CCC ATC CTC TGT GAC TC-3′, reverse: 5′-AGC TCA TAT GGG TCC GAC AG-3′); TNF-α (forward: 5′-ACG GCA TGG ATC TCA AAG AC-3′, reverse: 5′-AGA TAG CAA ATC GGC TGA CG-3′).

    Chloramphenicol Acetyltransferase Assay:

    Article Title: Erastin Inhibits Septic Shock and Inflammatory Gene Expression via Suppression of the NF-κB Pathway
    Article Snippet: Triplicate reactions were performed using the StepOnePlus Real-Time PCR (Thermo). .. Primer sequences were as follows: β-actin (forward: 5′- TTG CTG ACA GGA TGC AGA AG-3′, reverse: 5′-ACA TCT GCT GGA AGG TGG AC-3′); iNOS (forward: 5′-CAC CTT GGA GTT CAC CCA GT-3′, reverse: 5′-TGG TCA CCT CCA ACA CAA GA-3′); COX2 (forward: 5′- GGC CAT GGA GTG GAC TTA AA-3′, reverse: 5′-ACC TCT CCA CCA ATG ACC TG-3′); IL-1β (forward: 5′-GAG CCC ATC CTC TGT GAC TC-3′, reverse: 5′-AGC TCA TAT GGG TCC GAC AG-3′); TNF-α (forward: 5′-ACG GCA TGG ATC TCA AAG AC-3′, reverse: 5′-AGA TAG CAA ATC GGC TGA CG-3′).

    Titration:

    Article Title: Cross-Packaging and Capsid Mosaic Formation in Multiplexed AAV Libraries
    Article Snippet: Digested samples were then supplemented with 20 μL of 20 mg/mL proteinase K (740506; MACHEREY-NAGEL) and incubated at 70°C for 20 min. vgs were further extracted using the NucleoSpin RNA Virus kit (MACHEREY-NAGEL). qPCR was performed with a StepOnePlus Real-Time PCR (Life Technologies, Carlsbad, CA, USA). qPCR reactions were run in a final volume of 20 μL, containing the primers/probe PCR Master Mix (TaKaRa, Kusatsu, Japan) and 5 μL of template DNA. .. For ITR titration, the standard plasmid was prepared in accordance with D’Costa et al.

    Cellular Antioxidant Activity Assay:

    Article Title: Erastin Inhibits Septic Shock and Inflammatory Gene Expression via Suppression of the NF-κB Pathway
    Article Snippet: Triplicate reactions were performed using the StepOnePlus Real-Time PCR (Thermo). .. Primer sequences were as follows: β-actin (forward: 5′- TTG CTG ACA GGA TGC AGA AG-3′, reverse: 5′-ACA TCT GCT GGA AGG TGG AC-3′); iNOS (forward: 5′-CAC CTT GGA GTT CAC CCA GT-3′, reverse: 5′-TGG TCA CCT CCA ACA CAA GA-3′); COX2 (forward: 5′- GGC CAT GGA GTG GAC TTA AA-3′, reverse: 5′-ACC TCT CCA CCA ATG ACC TG-3′); IL-1β (forward: 5′-GAG CCC ATC CTC TGT GAC TC-3′, reverse: 5′-AGC TCA TAT GGG TCC GAC AG-3′); TNF-α (forward: 5′-ACG GCA TGG ATC TCA AAG AC-3′, reverse: 5′-AGA TAG CAA ATC GGC TGA CG-3′).

    Plasmid Preparation:

    Article Title: Cross-Packaging and Capsid Mosaic Formation in Multiplexed AAV Libraries
    Article Snippet: Digested samples were then supplemented with 20 μL of 20 mg/mL proteinase K (740506; MACHEREY-NAGEL) and incubated at 70°C for 20 min. vgs were further extracted using the NucleoSpin RNA Virus kit (MACHEREY-NAGEL). qPCR was performed with a StepOnePlus Real-Time PCR (Life Technologies, Carlsbad, CA, USA). qPCR reactions were run in a final volume of 20 μL, containing the primers/probe PCR Master Mix (TaKaRa, Kusatsu, Japan) and 5 μL of template DNA. .. For ITR titration, the standard plasmid was prepared in accordance with D’Costa et al.

    Reverse Transcription Polymerase Chain Reaction:

    Article Title: SOX9 expression decreases survival of patients with intrahepatic cholangiocarcinoma by conferring chemoresistance
    Article Snippet: Paragraph title: RNA isolation and quantitative real-time reverse transcription polymerase chain reaction ... For PCR amplification, 10.4 µL mixtures contained 5 µL (50 ng) template cDNA, 5 µL SYBR Green (4367659, Life Technologies) and 4 µM forward and reverse primer PCRs were run in triplicate and performed on a StepOnePlus Real-time PCR (Applied Biosystems).

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  • 99
    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: 99/100, 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 99 stars, based on 2976 article reviews
    Price from $9.99 to $1999.99
    real time pcr system - by Bioz Stars, 2020-04
    99/100 stars
      Buy from Supplier

    99
    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: 99/100, based on 3228 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/steponeplus real time system/product/Thermo Fisher
    Average 99 stars, based on 3228 article reviews
    Price from $9.99 to $1999.99
    steponeplus real time system - by Bioz Stars, 2020-04
    99/100 stars
      Buy from Supplier

    94
    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: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    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