fast sybr green master mix  (Thermo Fisher)


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    Name:
    Fast SYBR Green Master Mix
    Description:
    Alternative Product Try PowerUp SYBR Green Master Mix our newest high performance SYBR dye based master mix for superior performance at a very competitive price With PowerUp SYBR Green Master Mix we ve taken the best of Fast SYBR Green Master Mix and added additional capabilities for your gene expression analysis Fast SYBR Green Master Mix Real Time PCR Master Mix Designed for Speed Obtain a fast reliable and cost effective solution for your real time PCR applications without compromising sensitivity specificity dynamic range or PCR efficiency • Fast Real time PCR results in as fast as 35 minutes • Sensitive Detect very low copies of target • Specific Minimize primer dimer and non specific amplification • Reproducible Consistent amplification across a wide dynamic range Fast SYBR Green Master Mix contains all of the components excluding the template and primers in a convenient 2X master mix It includes the following components in an optimized buffer • AmpliTaq Fast DNA Polymerase UP a highly purified DNA polymerase designed to allow instant hot start minimizing non specific product formation and enabling reactions to be set up at room temperature • SYBR Green I dye to enable detection of double stranded DNA • Deoxynucleotides dNTPs to help maintain optimal PCR results • Uracil DNA Glycosylase UDG designed to reduce carryover contamination • Passive internal reference based on proprietary ROX dye to enable increased precision
    Catalog Number:
    4385610
    Price:
    None
    Applications:
    Enzymes & Master Mixes for Real-Time PCR|Fast Real-Time PCR|PCR & Real-Time PCR|Real Time PCR (qPCR)|Real Time PCR-Based Gene Expression Profiling|Gene Expression Analysis & Genotyping
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    Kits and Assays
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    Structured Review

    Thermo Fisher fast sybr green master mix
    HuR associates with SCN5A mRNA (A) Schematic representation of the SCN5A mRNA ARE sites in its 3′-UTR. The length of the mRNA is indicated by a kilo base (kb)-scale. (B) HuR binds to the SCN5A mRNA. After IP of RNA-protein complexes from cardiomyocytes using either anti-HuR antibody (Anti-HuR) or control IgG, RNA was isolated, and then, the cDNA was synthesized in a presence or absence of the reverse transcription enzyme. Two sets of primers to different regions of SCN5A were used to amplify SCN5A fragments. The PCR products were visualized with <t>SYBR</t> ™ Safe DNA Gel Stain in agarose gels. (C) <t>qRT-PCR</t> was performed to confirm the results in (B).
    Alternative Product Try PowerUp SYBR Green Master Mix our newest high performance SYBR dye based master mix for superior performance at a very competitive price With PowerUp SYBR Green Master Mix we ve taken the best of Fast SYBR Green Master Mix and added additional capabilities for your gene expression analysis Fast SYBR Green Master Mix Real Time PCR Master Mix Designed for Speed Obtain a fast reliable and cost effective solution for your real time PCR applications without compromising sensitivity specificity dynamic range or PCR efficiency • Fast Real time PCR results in as fast as 35 minutes • Sensitive Detect very low copies of target • Specific Minimize primer dimer and non specific amplification • Reproducible Consistent amplification across a wide dynamic range Fast SYBR Green Master Mix contains all of the components excluding the template and primers in a convenient 2X master mix It includes the following components in an optimized buffer • AmpliTaq Fast DNA Polymerase UP a highly purified DNA polymerase designed to allow instant hot start minimizing non specific product formation and enabling reactions to be set up at room temperature • SYBR Green I dye to enable detection of double stranded DNA • Deoxynucleotides dNTPs to help maintain optimal PCR results • Uracil DNA Glycosylase UDG designed to reduce carryover contamination • Passive internal reference based on proprietary ROX dye to enable increased precision
    https://www.bioz.com/result/fast sybr green master mix/product/Thermo Fisher
    Average 99 stars, based on 3358 article reviews
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    fast sybr green master mix - by Bioz Stars, 2020-07
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    Images

    1) Product Images from "HuR-mediated SCN5A mRNA stability reduces arrhythmic risk in heart failure"

    Article Title: HuR-mediated SCN5A mRNA stability reduces arrhythmic risk in heart failure

    Journal: Heart rhythm

    doi: 10.1016/j.hrthm.2018.02.018

    HuR associates with SCN5A mRNA (A) Schematic representation of the SCN5A mRNA ARE sites in its 3′-UTR. The length of the mRNA is indicated by a kilo base (kb)-scale. (B) HuR binds to the SCN5A mRNA. After IP of RNA-protein complexes from cardiomyocytes using either anti-HuR antibody (Anti-HuR) or control IgG, RNA was isolated, and then, the cDNA was synthesized in a presence or absence of the reverse transcription enzyme. Two sets of primers to different regions of SCN5A were used to amplify SCN5A fragments. The PCR products were visualized with SYBR ™ Safe DNA Gel Stain in agarose gels. (C) qRT-PCR was performed to confirm the results in (B).
    Figure Legend Snippet: HuR associates with SCN5A mRNA (A) Schematic representation of the SCN5A mRNA ARE sites in its 3′-UTR. The length of the mRNA is indicated by a kilo base (kb)-scale. (B) HuR binds to the SCN5A mRNA. After IP of RNA-protein complexes from cardiomyocytes using either anti-HuR antibody (Anti-HuR) or control IgG, RNA was isolated, and then, the cDNA was synthesized in a presence or absence of the reverse transcription enzyme. Two sets of primers to different regions of SCN5A were used to amplify SCN5A fragments. The PCR products were visualized with SYBR ™ Safe DNA Gel Stain in agarose gels. (C) qRT-PCR was performed to confirm the results in (B).

    Techniques Used: Isolation, Synthesized, Polymerase Chain Reaction, Staining, Quantitative RT-PCR

    2) Product Images from "Candida albicans Mycofilms Support Staphylococcus aureus Colonization and Enhances Miconazole Resistance in Dual-Species Interactions"

    Article Title: Candida albicans Mycofilms Support Staphylococcus aureus Colonization and Enhances Miconazole Resistance in Dual-Species Interactions

    Journal: Frontiers in Microbiology

    doi: 10.3389/fmicb.2017.00258

    Extracellular DNA contributes to inter-kingdom pathogenicity. Mono- and dual-species biofilms were seeded at 1 × 10 6 CFU/mL in black 96-well plates and eDNA release at 1.5, 6, 12, and 24 h measured using a SYBR ® Green 1 based microplate fluorescence assay (MFA) in comparison to a standard curve (A) . Biofilms were washed with 0.2 M EDTA to remove the ECM and resulting eDNA quantified using the MFA described above in comparison to a standard curve (B) . ECM associated DNA was then precipitated from matrix extracts and species contributions were analyzed using qPCR (C) . C. albicans only biofilms were grown for 24 h in black 96-well plates. After washing biofilms were then treated with either 130 or 650 mg/L of DNase for 4 h. SYTO9 ® stained S. aureus cells (1 × 10 6 CFU/mL) were then added to the biofilm and incubated for 90 mins before being fluorescently quantified in comparison to an vehicle control treated biofilm (D). Data represents duplicate samples from three independent experiments ( ∗ p
    Figure Legend Snippet: Extracellular DNA contributes to inter-kingdom pathogenicity. Mono- and dual-species biofilms were seeded at 1 × 10 6 CFU/mL in black 96-well plates and eDNA release at 1.5, 6, 12, and 24 h measured using a SYBR ® Green 1 based microplate fluorescence assay (MFA) in comparison to a standard curve (A) . Biofilms were washed with 0.2 M EDTA to remove the ECM and resulting eDNA quantified using the MFA described above in comparison to a standard curve (B) . ECM associated DNA was then precipitated from matrix extracts and species contributions were analyzed using qPCR (C) . C. albicans only biofilms were grown for 24 h in black 96-well plates. After washing biofilms were then treated with either 130 or 650 mg/L of DNase for 4 h. SYTO9 ® stained S. aureus cells (1 × 10 6 CFU/mL) were then added to the biofilm and incubated for 90 mins before being fluorescently quantified in comparison to an vehicle control treated biofilm (D). Data represents duplicate samples from three independent experiments ( ∗ p

    Techniques Used: SYBR Green Assay, Fluorescence, Real-time Polymerase Chain Reaction, Staining, Incubation

    3) Product Images from "Association of Impaired Reactive Aldehyde Metabolism with Delayed Graft Function in Human Kidney Transplantation"

    Article Title: Association of Impaired Reactive Aldehyde Metabolism with Delayed Graft Function in Human Kidney Transplantation

    Journal: Oxidative Medicine and Cellular Longevity

    doi: 10.1155/2018/3704129

    qPCR studies for kidney biopsies taken prior to transplant. For a separate series of kidney transplants, biopsies were collected for qPCR. (a) PCR primers used for qPCR. qPCR reactions were performed in a final volume of 20 μ l that contained 15 μ l of Fast SYBR© Green Master Mix (Life Technologies), 1000 nM primer (ALDH2, ALDH7A1 or GAPDH) or 500 nM primer (ALDH4A1), and 10 ng cDNA. (b–e) qPCR results for (b) ALDH2, (c) ALDH7A1, (d) ALDH4A1, and (e) ALDH1A1. ALDH2, ALDH7A1, and ALDH4A1 by qPCR were significantly elevated in kidneys that did not develop delayed graft function versus those kidneys that developed delayed graft function ( n = 10/group, ∗ P
    Figure Legend Snippet: qPCR studies for kidney biopsies taken prior to transplant. For a separate series of kidney transplants, biopsies were collected for qPCR. (a) PCR primers used for qPCR. qPCR reactions were performed in a final volume of 20 μ l that contained 15 μ l of Fast SYBR© Green Master Mix (Life Technologies), 1000 nM primer (ALDH2, ALDH7A1 or GAPDH) or 500 nM primer (ALDH4A1), and 10 ng cDNA. (b–e) qPCR results for (b) ALDH2, (c) ALDH7A1, (d) ALDH4A1, and (e) ALDH1A1. ALDH2, ALDH7A1, and ALDH4A1 by qPCR were significantly elevated in kidneys that did not develop delayed graft function versus those kidneys that developed delayed graft function ( n = 10/group, ∗ P

    Techniques Used: Real-time Polymerase Chain Reaction, Polymerase Chain Reaction, SYBR Green Assay

    4) Product Images from "A high-throughput qPCR system for simultaneous quantitative detection of dairy Lactococcus lactis and Leuconostoc bacteriophages"

    Article Title: A high-throughput qPCR system for simultaneous quantitative detection of dairy Lactococcus lactis and Leuconostoc bacteriophages

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0174223

    Performance of SYBR Green I and EvaGreen detection chemistries during qPCR assays. (A) Standard curves generated from amplification of serially diluted L . lactis phage P220 genome detected with the corresponding chemistries; (B) the corresponding performance parameters; and (C) amplification plots detected with SYBR Green I (brown) and EvaGreen (green).
    Figure Legend Snippet: Performance of SYBR Green I and EvaGreen detection chemistries during qPCR assays. (A) Standard curves generated from amplification of serially diluted L . lactis phage P220 genome detected with the corresponding chemistries; (B) the corresponding performance parameters; and (C) amplification plots detected with SYBR Green I (brown) and EvaGreen (green).

    Techniques Used: SYBR Green Assay, Real-time Polymerase Chain Reaction, Generated, Amplification

    5) Product Images from "Ufmylation and FATylation Pathways are Down Regulated in Human Alcoholic and Non Alcoholic Steatohepatitis, and Mice Fed DDC, where Mallory-Denk Bodies (MDBs) Form"

    Article Title: Ufmylation and FATylation Pathways are Down Regulated in Human Alcoholic and Non Alcoholic Steatohepatitis, and Mice Fed DDC, where Mallory-Denk Bodies (MDBs) Form

    Journal: Experimental and molecular pathology

    doi: 10.1016/j.yexmp.2014.05.010

    Induction of FAT10 and subunits of immunoproteasome (LMP2, LMP7 and MECL-1) in the livers of DDC re-fed mice. Quantification of mRNA was carried out by SYBR real-time PCR assays. n.s, not significant, * p
    Figure Legend Snippet: Induction of FAT10 and subunits of immunoproteasome (LMP2, LMP7 and MECL-1) in the livers of DDC re-fed mice. Quantification of mRNA was carried out by SYBR real-time PCR assays. n.s, not significant, * p

    Techniques Used: Mouse Assay, Real-time Polymerase Chain Reaction

    6) Product Images from "TRPV4 channel activation selectively inhibits tumor endothelial cell proliferation"

    Article Title: TRPV4 channel activation selectively inhibits tumor endothelial cell proliferation

    Journal: Scientific Reports

    doi: 10.1038/srep14257

    TEC express increased levels of proliferation-associated genes. Relative gene expression of cell-cycle associated genes in NEC and TEC. EC were treated with GSK (100 nM) for 24 h and subsequently lysed for RNA isolation. cDNA was prepared and qPCR analysis was performed using Fast SYBR green master mix (Applied Biosystems). Gene expression was first normalized to GAPDH and presented as relative expression to NEC. All the data shown is mean ± SEM from at least three independent experiments.
    Figure Legend Snippet: TEC express increased levels of proliferation-associated genes. Relative gene expression of cell-cycle associated genes in NEC and TEC. EC were treated with GSK (100 nM) for 24 h and subsequently lysed for RNA isolation. cDNA was prepared and qPCR analysis was performed using Fast SYBR green master mix (Applied Biosystems). Gene expression was first normalized to GAPDH and presented as relative expression to NEC. All the data shown is mean ± SEM from at least three independent experiments.

    Techniques Used: Expressing, Isolation, Real-time Polymerase Chain Reaction, SYBR Green Assay

    7) Product Images from "Development of a Sensitive Real-Time Fast-qPCR Based on SYBR® Green for Detection and Quantification of Chicken Parvovirus (ChPV)"

    Article Title: Development of a Sensitive Real-Time Fast-qPCR Based on SYBR® Green for Detection and Quantification of Chicken Parvovirus (ChPV)

    Journal: Veterinary Sciences

    doi: 10.3390/vetsci5030069

    Detection and quantitation of ChPV using the sensitive fast-qPCR based on SYBR ® Green. ChPV = Chicken Parvovirus; BC = Broiler Chickens; LH = Layer Hens; BH = Breeder Hens; CG = Copies of Genome.
    Figure Legend Snippet: Detection and quantitation of ChPV using the sensitive fast-qPCR based on SYBR ® Green. ChPV = Chicken Parvovirus; BC = Broiler Chickens; LH = Layer Hens; BH = Breeder Hens; CG = Copies of Genome.

    Techniques Used: Quantitation Assay, Real-time Polymerase Chain Reaction, SYBR Green Assay

    Sensitive real time fast-real time PCR (qPCR) based on SYBR ® Green for detection and quantification of non-structural (NS) gene of Chicken Parvovirus (ChPV)—( A ) Standard curve using 10-fold serial dilution of NS gene plasmid of ChPV; ( B ) Amplification Plot; ( C ) Melting Curve.
    Figure Legend Snippet: Sensitive real time fast-real time PCR (qPCR) based on SYBR ® Green for detection and quantification of non-structural (NS) gene of Chicken Parvovirus (ChPV)—( A ) Standard curve using 10-fold serial dilution of NS gene plasmid of ChPV; ( B ) Amplification Plot; ( C ) Melting Curve.

    Techniques Used: Real-time Polymerase Chain Reaction, SYBR Green Assay, Serial Dilution, Plasmid Preparation, Amplification

    8) Product Images from "Central Resistin Overexposure Induces Insulin Resistance Through Toll-Like Receptor 4"

    Article Title: Central Resistin Overexposure Induces Insulin Resistance Through Toll-Like Receptor 4

    Journal: Diabetes

    doi: 10.2337/db12-0237

    Effect of chronic resistin treatment on SOCS-3, PTP-1B, and IL-6 expression. Male Wistar rats received an ICV of vehicle or resistin (1.2 μg/day) during a period of 2 weeks. At the end of the infusion period, SOCS-3 ( A ), PTP-1B ( B ), and IL-6 ( C ) mRNA levels were evaluated by SYBR Green real-time RT-PCR (Applied Biosystems) in the hypothalamus and peripheral insulin-sensitive tissues. In SH-SY5Y cells, mRNA levels of SOCS-3 and PTP-1B were measured after chronic resistin treatment (100 ng/mL) for 16 h. Results were normalized to 18S RNA. For protein analysis, protein lysates from hypothalamus, liver, muscle, adipose tissue, and SH-SY5Y cells were subjected to Western blot analysis. Membranes were probed sequentially with anti–SOCS-3 ( A ), anti–PTP-1B ( B ), and anti–IL-6 ( C ). The proteins on the blots were revealed by ECL and bands quantified by densitometry. The results are expressed as ratio of SOCS-3/β-tubulin, PTP-1B/β-tubulin, and IL-6/β-tubulin. Data are means ± SEM ( n = 3–6/group). * P
    Figure Legend Snippet: Effect of chronic resistin treatment on SOCS-3, PTP-1B, and IL-6 expression. Male Wistar rats received an ICV of vehicle or resistin (1.2 μg/day) during a period of 2 weeks. At the end of the infusion period, SOCS-3 ( A ), PTP-1B ( B ), and IL-6 ( C ) mRNA levels were evaluated by SYBR Green real-time RT-PCR (Applied Biosystems) in the hypothalamus and peripheral insulin-sensitive tissues. In SH-SY5Y cells, mRNA levels of SOCS-3 and PTP-1B were measured after chronic resistin treatment (100 ng/mL) for 16 h. Results were normalized to 18S RNA. For protein analysis, protein lysates from hypothalamus, liver, muscle, adipose tissue, and SH-SY5Y cells were subjected to Western blot analysis. Membranes were probed sequentially with anti–SOCS-3 ( A ), anti–PTP-1B ( B ), and anti–IL-6 ( C ). The proteins on the blots were revealed by ECL and bands quantified by densitometry. The results are expressed as ratio of SOCS-3/β-tubulin, PTP-1B/β-tubulin, and IL-6/β-tubulin. Data are means ± SEM ( n = 3–6/group). * P

    Techniques Used: Expressing, SYBR Green Assay, Quantitative RT-PCR, Western Blot

    Effect of central chronic resistin infusion on metabolic and endocrine parameters. A – C : Male Wistar rats received an ICV of vehicle or resistin (1.2 μg/day) during a period of 2 weeks, and body weight gain and food intake were measured daily ( A ). At the end of the infusion period, the mRNA levels of POMC and NPY were measured in the hypothalamus ( B ). UCP-2, adipoR1, and adipoR2 mRNA levels also measured in the hypothalamus and peripheral insulin-sensitive tissues using SYBR Green real-time RT-PCR (Applied Biosystems). Results were normalized to 18S RNA. All data are presented as means ± SEM ( n = 3–6/group). * P
    Figure Legend Snippet: Effect of central chronic resistin infusion on metabolic and endocrine parameters. A – C : Male Wistar rats received an ICV of vehicle or resistin (1.2 μg/day) during a period of 2 weeks, and body weight gain and food intake were measured daily ( A ). At the end of the infusion period, the mRNA levels of POMC and NPY were measured in the hypothalamus ( B ). UCP-2, adipoR1, and adipoR2 mRNA levels also measured in the hypothalamus and peripheral insulin-sensitive tissues using SYBR Green real-time RT-PCR (Applied Biosystems). Results were normalized to 18S RNA. All data are presented as means ± SEM ( n = 3–6/group). * P

    Techniques Used: SYBR Green Assay, Quantitative RT-PCR

    Effect of chronic resistin treatment on IR phosphorylation and expression. Male Wistar rats received an ICV of vehicle or resistin (1.2 μg/day) during a period of 2 weeks. At the end of the infusion period, vehicle and resistin-treated rats received IP human insulin (1 U/kg body weight) or saline bolus 30 min before euthanasia to test insulin sensitivity. Serum-deprived SH-SY5Y cells were incubated for 16 h with resistin (100 ng/mL) and then stimulated for 10 min with or without insulin (100 nmol/L). Protein lysates from hypothalamus, liver, muscle, adipose tissue, and SH-SY5Y cells were subjected to Western blot analysis. Membranes were probed sequentially with anti– p -IR ( A ) followed by anti-IR antibodies ( B ). The proteins on the blots were revealed by ECL and bands quantified by densitometry. The results are expressed as ratio of p -IR/β-tubulin and IR/β-tubulin. C : At the end of the experimental period of ICV resistin, SYBR Green real-time RT-PCR (Applied Biosystems) was conducted to measure IR mRNA levels in the hypothalamus and peripheral insulin-sensitive tissues. In SH-SY5Y cells, IR mRNA expression was measured after chronic resistin treatment (100 ng/mL) for 16 h. Results were normalized to 18S RNA. All data are means ± SEM ( n = 3–6/group). a,b,c denote significant differences at P
    Figure Legend Snippet: Effect of chronic resistin treatment on IR phosphorylation and expression. Male Wistar rats received an ICV of vehicle or resistin (1.2 μg/day) during a period of 2 weeks. At the end of the infusion period, vehicle and resistin-treated rats received IP human insulin (1 U/kg body weight) or saline bolus 30 min before euthanasia to test insulin sensitivity. Serum-deprived SH-SY5Y cells were incubated for 16 h with resistin (100 ng/mL) and then stimulated for 10 min with or without insulin (100 nmol/L). Protein lysates from hypothalamus, liver, muscle, adipose tissue, and SH-SY5Y cells were subjected to Western blot analysis. Membranes were probed sequentially with anti– p -IR ( A ) followed by anti-IR antibodies ( B ). The proteins on the blots were revealed by ECL and bands quantified by densitometry. The results are expressed as ratio of p -IR/β-tubulin and IR/β-tubulin. C : At the end of the experimental period of ICV resistin, SYBR Green real-time RT-PCR (Applied Biosystems) was conducted to measure IR mRNA levels in the hypothalamus and peripheral insulin-sensitive tissues. In SH-SY5Y cells, IR mRNA expression was measured after chronic resistin treatment (100 ng/mL) for 16 h. Results were normalized to 18S RNA. All data are means ± SEM ( n = 3–6/group). a,b,c denote significant differences at P

    Techniques Used: Expressing, Incubation, Western Blot, SYBR Green Assay, Quantitative RT-PCR

    9) Product Images from "Association of Impaired Reactive Aldehyde Metabolism with Delayed Graft Function in Human Kidney Transplantation"

    Article Title: Association of Impaired Reactive Aldehyde Metabolism with Delayed Graft Function in Human Kidney Transplantation

    Journal: Oxidative Medicine and Cellular Longevity

    doi: 10.1155/2018/3704129

    qPCR studies for kidney biopsies taken prior to transplant. For a separate series of kidney transplants, biopsies were collected for qPCR. (a) PCR primers used for qPCR. qPCR reactions were performed in a final volume of 20 μ l that contained 15 μ l of Fast SYBR© Green Master Mix (Life Technologies), 1000 nM primer (ALDH2, ALDH7A1 or GAPDH) or 500 nM primer (ALDH4A1), and 10 ng cDNA. (b–e) qPCR results for (b) ALDH2, (c) ALDH7A1, (d) ALDH4A1, and (e) ALDH1A1. ALDH2, ALDH7A1, and ALDH4A1 by qPCR were significantly elevated in kidneys that did not develop delayed graft function versus those kidneys that developed delayed graft function ( n = 10/group, ∗ P
    Figure Legend Snippet: qPCR studies for kidney biopsies taken prior to transplant. For a separate series of kidney transplants, biopsies were collected for qPCR. (a) PCR primers used for qPCR. qPCR reactions were performed in a final volume of 20 μ l that contained 15 μ l of Fast SYBR© Green Master Mix (Life Technologies), 1000 nM primer (ALDH2, ALDH7A1 or GAPDH) or 500 nM primer (ALDH4A1), and 10 ng cDNA. (b–e) qPCR results for (b) ALDH2, (c) ALDH7A1, (d) ALDH4A1, and (e) ALDH1A1. ALDH2, ALDH7A1, and ALDH4A1 by qPCR were significantly elevated in kidneys that did not develop delayed graft function versus those kidneys that developed delayed graft function ( n = 10/group, ∗ P

    Techniques Used: Real-time Polymerase Chain Reaction, Polymerase Chain Reaction, SYBR Green Assay

    10) Product Images from "Long Interleukin-22 Binding Protein Isoform-1 Is an Intracellular Activator of the Unfolded Protein Response"

    Article Title: Long Interleukin-22 Binding Protein Isoform-1 Is an Intracellular Activator of the Unfolded Protein Response

    Journal: Frontiers in Immunology

    doi: 10.3389/fimmu.2018.02934

    IL-22BP protein and mRNA levels in monocyte-derived DCs. (A) (Upper left) Diagram showing exons with the primer-binding positions for IL22RA2 variant discrimination strategy. (Lower left) Expression of IL22RA2v1, v2 , and v3 in moDC after 6 days of culture in differentiation medium (DM) ± AM580, a retinoic acid receptor agonist, by RT-PCR. IL22RA2v2 expression vector was used as positive control for IL22RA2v2 expression. (Upper right) Expression of IL22RA2 variants in moDC following 6 days of culture in DM ± AM580 by RT-qPCR using the isoform-specific primers indicated in the left diagram (mean ± SEM; n = 3) relative to the housekeeping gene GAPDH . (B) Effect of different maturation stimuli (CpG and LPS + IFN-γ or Poly I:C) on IL22RA2, IL12B , and IL6 expression in moDC after 6 days of culture in DM by RT-qPCR using Taqman probes relative to the housekeeping gene ACTB . Extent of maturation is represented as percentage of CD83 + fluorescence by flow cytometry. Data showing representative experiment of 3 performed. (C) IL-22BP secretion by moDC cultured in DM detected by ELISA corresponds to IL22RA2 mRNA expression levels analyzed by RT-qPCR using Taqman probes and both increase over the cultivation period (mean ± SEM, n = 2). (D) Cell lysates (CL) of moDCs and conditioned media (CM) were harvested after 6 days in DM ± AM580 and immunoblotted for detection of IL-22BP (IL-22BP antibody 1) using tubulin as a loading control, IL22RA2 mRNA expression relative to the mean of the housekeeping genes GAPDH and ACTB was measured by RT-qPCR using pre-designed SYBR Green primers from the same cells. Representative experiment out of three performed. All primers used are listed in Supplementary Table 2 .
    Figure Legend Snippet: IL-22BP protein and mRNA levels in monocyte-derived DCs. (A) (Upper left) Diagram showing exons with the primer-binding positions for IL22RA2 variant discrimination strategy. (Lower left) Expression of IL22RA2v1, v2 , and v3 in moDC after 6 days of culture in differentiation medium (DM) ± AM580, a retinoic acid receptor agonist, by RT-PCR. IL22RA2v2 expression vector was used as positive control for IL22RA2v2 expression. (Upper right) Expression of IL22RA2 variants in moDC following 6 days of culture in DM ± AM580 by RT-qPCR using the isoform-specific primers indicated in the left diagram (mean ± SEM; n = 3) relative to the housekeeping gene GAPDH . (B) Effect of different maturation stimuli (CpG and LPS + IFN-γ or Poly I:C) on IL22RA2, IL12B , and IL6 expression in moDC after 6 days of culture in DM by RT-qPCR using Taqman probes relative to the housekeeping gene ACTB . Extent of maturation is represented as percentage of CD83 + fluorescence by flow cytometry. Data showing representative experiment of 3 performed. (C) IL-22BP secretion by moDC cultured in DM detected by ELISA corresponds to IL22RA2 mRNA expression levels analyzed by RT-qPCR using Taqman probes and both increase over the cultivation period (mean ± SEM, n = 2). (D) Cell lysates (CL) of moDCs and conditioned media (CM) were harvested after 6 days in DM ± AM580 and immunoblotted for detection of IL-22BP (IL-22BP antibody 1) using tubulin as a loading control, IL22RA2 mRNA expression relative to the mean of the housekeeping genes GAPDH and ACTB was measured by RT-qPCR using pre-designed SYBR Green primers from the same cells. Representative experiment out of three performed. All primers used are listed in Supplementary Table 2 .

    Techniques Used: Derivative Assay, Binding Assay, Variant Assay, Expressing, Reverse Transcription Polymerase Chain Reaction, Plasmid Preparation, Positive Control, Quantitative RT-PCR, Fluorescence, Flow Cytometry, Cytometry, Cell Culture, Enzyme-linked Immunosorbent Assay, SYBR Green Assay

    11) Product Images from "TLR3/4 Signaling is Mediated via the NFκB-CXCR4/7 Pathway in Human Alcoholic Hepatitis and Non Alcoholic Steatohepatitis Which Formed Mallory-Denk Bodies"

    Article Title: TLR3/4 Signaling is Mediated via the NFκB-CXCR4/7 Pathway in Human Alcoholic Hepatitis and Non Alcoholic Steatohepatitis Which Formed Mallory-Denk Bodies

    Journal: Experimental and molecular pathology

    doi: 10.1016/j.yexmp.2014.07.001

    Induction of TLRs downstream components in the livers of AH (A) and NASH (B) biopsies. Quantification of mRNA was carried out by SYBR real-time PCR assays. * p
    Figure Legend Snippet: Induction of TLRs downstream components in the livers of AH (A) and NASH (B) biopsies. Quantification of mRNA was carried out by SYBR real-time PCR assays. * p

    Techniques Used: Real-time Polymerase Chain Reaction

    12) Product Images from "Unusual Five Copies and Dual Forms of nrdB in “Candidatus Liberibacter asiaticus”: Biological Implications and PCR Detection Application"

    Article Title: Unusual Five Copies and Dual Forms of nrdB in “Candidatus Liberibacter asiaticus”: Biological Implications and PCR Detection Application

    Journal: Scientific Reports

    doi: 10.1038/srep39020

    Comparisons of PCR detection sensitivities on “ Candidatus Liberibacter asiaticus” using 57 samples from China (34) and USA (23) among primer sets RNRf/RNRr ( nrdB -based), HLBas/HLBr (16S rRNA gene-based), and LJ900f/LJ900r (prophage-based). ( a ) SYBR Green real-time PCR. ( b ) TaqMan Real-time PCR. Numbers within each bar box are mean Ct values with standard deviation. P values were calculated based on independent-sample T-test. All qPCR assays were performed on the ABI real-time PCR system with the same regent kit (Universal PCR Master Mix, Applied biosystems).
    Figure Legend Snippet: Comparisons of PCR detection sensitivities on “ Candidatus Liberibacter asiaticus” using 57 samples from China (34) and USA (23) among primer sets RNRf/RNRr ( nrdB -based), HLBas/HLBr (16S rRNA gene-based), and LJ900f/LJ900r (prophage-based). ( a ) SYBR Green real-time PCR. ( b ) TaqMan Real-time PCR. Numbers within each bar box are mean Ct values with standard deviation. P values were calculated based on independent-sample T-test. All qPCR assays were performed on the ABI real-time PCR system with the same regent kit (Universal PCR Master Mix, Applied biosystems).

    Techniques Used: Polymerase Chain Reaction, SYBR Green Assay, Real-time Polymerase Chain Reaction, Standard Deviation

    13) Product Images from "Development of a Sensitive Real-Time Fast-qPCR Based on SYBR® Green for Detection and Quantification of Chicken Parvovirus (ChPV)"

    Article Title: Development of a Sensitive Real-Time Fast-qPCR Based on SYBR® Green for Detection and Quantification of Chicken Parvovirus (ChPV)

    Journal: Veterinary Sciences

    doi: 10.3390/vetsci5030069

    Sensitive real time fast-real time PCR (qPCR) based on SYBR ® Green for detection and quantification of non-structural (NS) gene of Chicken Parvovirus (ChPV)—( A ) Standard curve using 10-fold serial dilution of NS gene plasmid of ChPV; ( B ) Amplification Plot; ( C ) Melting Curve.
    Figure Legend Snippet: Sensitive real time fast-real time PCR (qPCR) based on SYBR ® Green for detection and quantification of non-structural (NS) gene of Chicken Parvovirus (ChPV)—( A ) Standard curve using 10-fold serial dilution of NS gene plasmid of ChPV; ( B ) Amplification Plot; ( C ) Melting Curve.

    Techniques Used: Real-time Polymerase Chain Reaction, SYBR Green Assay, Serial Dilution, Plasmid Preparation, Amplification

    14) Product Images from "RBCS1A and RBCS3B, two major members within the Arabidopsis RBCS multigene family, function to yield sufficient Rubisco content for leaf photosynthetic capacity"

    Article Title: RBCS1A and RBCS3B, two major members within the Arabidopsis RBCS multigene family, function to yield sufficient Rubisco content for leaf photosynthetic capacity

    Journal: Journal of Experimental Botany

    doi: 10.1093/jxb/err434

    Identification of rbcs1a-1, rbcs3b-1 , and rbcs1a3b-1 mutants. (A) Genomic structures of RBCS1A and RBCS3B loci and T-DNA insertion sites. White and black boxes represent exons. The white boxes represent the untranslated regions and the black boxes represent the coding regions in exons. Grey boxes represent T-DNA. Arrows represent the positions of primer pairs used for RT-PCR analysis. (B) RT-PCR analysis for investigating RBCS1A and RBCS3B mRNA accumulation in leaves of rbcs mutants. Total RNA was isolated from leaves of wild-type, rbcs1a-1, rbcs3b-1 , and rbcs1a3b-1 plants and subjected to RT-PCR analysis using gene-specific primers. PCR products were separated by electrophoresis, stained with SYBR Green I, and detected by a fluorescence image analyser. 18S rRNA was used as an internal control.
    Figure Legend Snippet: Identification of rbcs1a-1, rbcs3b-1 , and rbcs1a3b-1 mutants. (A) Genomic structures of RBCS1A and RBCS3B loci and T-DNA insertion sites. White and black boxes represent exons. The white boxes represent the untranslated regions and the black boxes represent the coding regions in exons. Grey boxes represent T-DNA. Arrows represent the positions of primer pairs used for RT-PCR analysis. (B) RT-PCR analysis for investigating RBCS1A and RBCS3B mRNA accumulation in leaves of rbcs mutants. Total RNA was isolated from leaves of wild-type, rbcs1a-1, rbcs3b-1 , and rbcs1a3b-1 plants and subjected to RT-PCR analysis using gene-specific primers. PCR products were separated by electrophoresis, stained with SYBR Green I, and detected by a fluorescence image analyser. 18S rRNA was used as an internal control.

    Techniques Used: Reverse Transcription Polymerase Chain Reaction, Isolation, Polymerase Chain Reaction, Electrophoresis, Staining, SYBR Green Assay, Fluorescence

    15) Product Images from "Single-Cell Analysis of Murine Long-Term Hematopoietic Stem Cells Reveals Distinct Patterns of Gene Expression during Fetal Migration"

    Article Title: Single-Cell Analysis of Murine Long-Term Hematopoietic Stem Cells Reveals Distinct Patterns of Gene Expression during Fetal Migration

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0030542

    Validation of single-cell multiplex RT-qPCR. A) Schematic representation of the single cell multiplex RT-qPCR methodology. After lysing single cells by heating, cDNA is obtained by retrotranscription with as many specific primers as genes studied. The cDNA template is subjected to a 10-cycle pre-amplification reaction using a primer pair for each gene studied in order to maintain the linear ratio of the original sample. The pre-amplified product is split into aliquots for qPCR analysis with SYBR®-Green using the same primer pairs as in the pre-amplification step. B) Efficiency of qPCR reactions after pre-amplification using 10-fold dilutions of standards did not show significant differences between the genes studied. C) Similar qPCR Ct values between samples using either single or multiplex RT primers indicate no primer competition during the retrotranscription reaction. D) Similar qPCR Ct values between samples using either single or multiplex primer pairs indicate no primer competition during the 10-cycle pre-amplification reaction. E) Quantification of varied copy number for a particular gene amplified in the presence of a constant number of plasmids for the other genes studied. For each reaction, 10 2 (purple line), 10 4 (blue line), or 10 6 (red line) plasmids were mixed with a constant number (10 4 ) of plasmids for the other eight genes studied. Gene specific multiplex qPCR amplification was then performed. Each reaction was performed in triplicate. F) Regression lines obtained from standard curves for each gene with the single cell RT-qPCR conditions. R 2 -values were greater than 0.990 with detection limit of 100 copies for most genes.
    Figure Legend Snippet: Validation of single-cell multiplex RT-qPCR. A) Schematic representation of the single cell multiplex RT-qPCR methodology. After lysing single cells by heating, cDNA is obtained by retrotranscription with as many specific primers as genes studied. The cDNA template is subjected to a 10-cycle pre-amplification reaction using a primer pair for each gene studied in order to maintain the linear ratio of the original sample. The pre-amplified product is split into aliquots for qPCR analysis with SYBR®-Green using the same primer pairs as in the pre-amplification step. B) Efficiency of qPCR reactions after pre-amplification using 10-fold dilutions of standards did not show significant differences between the genes studied. C) Similar qPCR Ct values between samples using either single or multiplex RT primers indicate no primer competition during the retrotranscription reaction. D) Similar qPCR Ct values between samples using either single or multiplex primer pairs indicate no primer competition during the 10-cycle pre-amplification reaction. E) Quantification of varied copy number for a particular gene amplified in the presence of a constant number of plasmids for the other genes studied. For each reaction, 10 2 (purple line), 10 4 (blue line), or 10 6 (red line) plasmids were mixed with a constant number (10 4 ) of plasmids for the other eight genes studied. Gene specific multiplex qPCR amplification was then performed. Each reaction was performed in triplicate. F) Regression lines obtained from standard curves for each gene with the single cell RT-qPCR conditions. R 2 -values were greater than 0.990 with detection limit of 100 copies for most genes.

    Techniques Used: Multiplex Assay, Quantitative RT-PCR, Amplification, Real-time Polymerase Chain Reaction, SYBR Green Assay

    Related Articles

    SYBR Green Assay:

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    Article Snippet: .. For qRT-PCR Fast SYBR Green Master Mix (Applied Biosystems) as per manufacturer’s instruction and the StepOnePlus Real-Time PCR system (Applied Biosystems) were used. .. Fast SYBR Green Master Mix contains the fluorescent ROX as a passive reference.

    Article Title: Candida albicans Mycofilms Support Staphylococcus aureus Colonization and Enhances Miconazole Resistance in Dual-Species Interactions
    Article Snippet: .. Briefly, 1 μL of extracted DNA was added to a PCR mastermix containing Fast SYBR® Green Master Mix, 10 μM species-specific forward and reverse primers ( Table ), and RNase free water. qPCR was then carried out using the Step-One plus real time PCR machine (Life Technologies, Paisley, UK) using the following thermal profile: 50°C for 2 min, 95°C for 2 min, followed by 40 cycles of 95°C for 3 s and 60°C for 30 s. Colony forming equivalents (CFE) were calculated compared to a standard curve of serially diluted DNA of each species as previously described ( ). .. Dual-Species Biofilm Visualization Mono- and dual-species biofilms were stained with 5 μM calcofluor white (Invitrogen, Paisley, UK) and 20 μM SYTO9® (Sigma–Aldrich, Dorset, UK) was used to stain both fungal and bacterial cells.

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    Article Title: Ovarian carcinomas with genetic and epigenetic BRCA1 loss have distinct molecular abnormalities
    Article Snippet: .. Samples (10 ng bisulfite-treated DNA) were run in triplicate containing 5 μL SYBR Green Master Mix (Applied Biosystems, Foster City, CA) and 5 pmol of each forward and reverse primer. .. Bisulfite-converted CpG methylated Jurkat Genomic DNA (New England Biolabs, Ipswich, MA) served as a positive control and was used to generate a standard curve to quantify the amount of fully methylated promoters in each reaction.

    Article Title: HuR-mediated SCN5A mRNA stability reduces arrhythmic risk in heart failure
    Article Snippet: .. Quantitative real-time reverse-transcriptase polymerase chain reaction (qRT-PCR) was carried out using gene-specific primers, Fast SYBR® Green Master Mix and 7500 Fast Real-Time PCR System (Thermo Fisher Scientific, Waltham, MA). .. Gene expression levels were normalized to the level of β-actin.

    Real-time Polymerase Chain Reaction:

    Article Title: Signaling Properties and Pharmacological Analysis of Two Sulfakinin Receptors from the Red Flour Beetle, Tribolium castaneum
    Article Snippet: .. For qRT-PCR Fast SYBR Green Master Mix (Applied Biosystems) as per manufacturer’s instruction and the StepOnePlus Real-Time PCR system (Applied Biosystems) were used. .. Fast SYBR Green Master Mix contains the fluorescent ROX as a passive reference.

    Article Title: Candida albicans Mycofilms Support Staphylococcus aureus Colonization and Enhances Miconazole Resistance in Dual-Species Interactions
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    Article Title: Association of Impaired Reactive Aldehyde Metabolism with Delayed Graft Function in Human Kidney Transplantation
    Article Snippet: .. To substantially reduce the possibility of DNA contamination in the preparations, the isolated total RNA was subject to precipitation with lithium chloride and DNase digestion (Ambion DNase-free). cDNA was made using the Takara© Primescript cDNA synthesis kit with oligo dT primers. qPCR reactions were performed in a final volume of 20 μ l that contained 15 μ l of Fast SYBR© Green Master Mix (Life Technologies), 1000 nM primer (ALDH2, ALDH7A1, or GAPDH) or 500 nM primer (ALDH4A1), and 10 ng cDNA. ..

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    Article Title: HuR-mediated SCN5A mRNA stability reduces arrhythmic risk in heart failure
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    Polymerase Chain Reaction:

    Article Title: Candida albicans Mycofilms Support Staphylococcus aureus Colonization and Enhances Miconazole Resistance in Dual-Species Interactions
    Article Snippet: .. Briefly, 1 μL of extracted DNA was added to a PCR mastermix containing Fast SYBR® Green Master Mix, 10 μM species-specific forward and reverse primers ( Table ), and RNase free water. qPCR was then carried out using the Step-One plus real time PCR machine (Life Technologies, Paisley, UK) using the following thermal profile: 50°C for 2 min, 95°C for 2 min, followed by 40 cycles of 95°C for 3 s and 60°C for 30 s. Colony forming equivalents (CFE) were calculated compared to a standard curve of serially diluted DNA of each species as previously described ( ). .. Dual-Species Biofilm Visualization Mono- and dual-species biofilms were stained with 5 μM calcofluor white (Invitrogen, Paisley, UK) and 20 μM SYTO9® (Sigma–Aldrich, Dorset, UK) was used to stain both fungal and bacterial cells.

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    Article Title: HuR-mediated SCN5A mRNA stability reduces arrhythmic risk in heart failure
    Article Snippet: .. Quantitative real-time reverse-transcriptase polymerase chain reaction (qRT-PCR) was carried out using gene-specific primers, Fast SYBR® Green Master Mix and 7500 Fast Real-Time PCR System (Thermo Fisher Scientific, Waltham, MA). .. Gene expression levels were normalized to the level of β-actin.

    Quantitative RT-PCR:

    Article Title: Signaling Properties and Pharmacological Analysis of Two Sulfakinin Receptors from the Red Flour Beetle, Tribolium castaneum
    Article Snippet: .. For qRT-PCR Fast SYBR Green Master Mix (Applied Biosystems) as per manufacturer’s instruction and the StepOnePlus Real-Time PCR system (Applied Biosystems) were used. .. Fast SYBR Green Master Mix contains the fluorescent ROX as a passive reference.

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    Isolation:

    Article Title: Association of Impaired Reactive Aldehyde Metabolism with Delayed Graft Function in Human Kidney Transplantation
    Article Snippet: .. To substantially reduce the possibility of DNA contamination in the preparations, the isolated total RNA was subject to precipitation with lithium chloride and DNase digestion (Ambion DNase-free). cDNA was made using the Takara© Primescript cDNA synthesis kit with oligo dT primers. qPCR reactions were performed in a final volume of 20 μ l that contained 15 μ l of Fast SYBR© Green Master Mix (Life Technologies), 1000 nM primer (ALDH2, ALDH7A1, or GAPDH) or 500 nM primer (ALDH4A1), and 10 ng cDNA. ..

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    Thermo Fisher fast sybr green master mix
    HuR associates with SCN5A mRNA (A) Schematic representation of the SCN5A mRNA ARE sites in its 3′-UTR. The length of the mRNA is indicated by a kilo base (kb)-scale. (B) HuR binds to the SCN5A mRNA. After IP of RNA-protein complexes from cardiomyocytes using either anti-HuR antibody (Anti-HuR) or control IgG, RNA was isolated, and then, the cDNA was synthesized in a presence or absence of the reverse transcription enzyme. Two sets of primers to different regions of SCN5A were used to amplify SCN5A fragments. The PCR products were visualized with <t>SYBR</t> ™ Safe DNA Gel Stain in agarose gels. (C) <t>qRT-PCR</t> was performed to confirm the results in (B).
    Fast Sybr Green Master Mix, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 3358 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    84
    Thermo Fisher taqman protein assays fast master mix
    Effect of variant region deletion on 3D structure and gene expression (A) Combined 3C-qPCR data of chromatin interactions in dVR cells. Black: control; Blue: dVR. The numbering of NcoI DNA fragments is given relative to the viewpoint. Viewing point: R13 region; Black bars: STARD10 exon; red bars: regulatory region; orange box: qPCR probe; orange arrow: qPCR constant primer; green stars: credible set genetic variants. Data were normalized to a CXCL12 loading control. n = 3. (B) <t>Taqman</t> <t>qRT-PCR</t> analysis of gene expression in control and dVR cells. *, P
    Taqman Protein Assays Fast Master Mix, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 84/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher fast universal pcr master mix
    Analysis of BMP4 and CST6 mRNA levels following treatment with demethylating agent and histone deacetylase inhibitor. (A) A CpG island (506 bp) spans the transcription start site of the human BMP4 gene. (B) Relative BMP4 mRNA levels in vehicle (DMSO)-treated 231_ATCC and 231_HM.LNm5 cells by <t>TaqMan</t> <t>qRT-PCR.</t> Expression in 231_ATCC was set to 1. Mean±s.d. ( n =3). (C) TaqMan qRT-PCR analysis of BMP4 mRNA expression levels in 5-Aza-2′-deoxycytidine (5azadC)- and Trichostatin A (TSA)-treated 231_ATCC and 231_HM.LNm5 cells. Expression in vehicle-treated cells was set to 1. Mean±s.d. ( n =3). (D) A CpG island (370 bp) spans the transcription start site of the human CST6 gene. (E) Relative CST6 mRNA levels in vehicle (DMSO)-treated 231_ATCC and 231_HM.LNm5 cells by TaqMan qRT-PCR. Expression in 231_ATCC was set to 1. Mean±s.d. ( n =3). (F) TaqMan qRT-PCR analysis of CST6 mRNA expression levels in 5azadC- and TSA-treated 231_ATCC and 231_HM.LNm5 cells. Expression in vehicle-treated cells was set to 1. Mean±s.d. ( n =3).
    Fast Universal Pcr Master Mix, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 32 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    HuR associates with SCN5A mRNA (A) Schematic representation of the SCN5A mRNA ARE sites in its 3′-UTR. The length of the mRNA is indicated by a kilo base (kb)-scale. (B) HuR binds to the SCN5A mRNA. After IP of RNA-protein complexes from cardiomyocytes using either anti-HuR antibody (Anti-HuR) or control IgG, RNA was isolated, and then, the cDNA was synthesized in a presence or absence of the reverse transcription enzyme. Two sets of primers to different regions of SCN5A were used to amplify SCN5A fragments. The PCR products were visualized with SYBR ™ Safe DNA Gel Stain in agarose gels. (C) qRT-PCR was performed to confirm the results in (B).

    Journal: Heart rhythm

    Article Title: HuR-mediated SCN5A mRNA stability reduces arrhythmic risk in heart failure

    doi: 10.1016/j.hrthm.2018.02.018

    Figure Lengend Snippet: HuR associates with SCN5A mRNA (A) Schematic representation of the SCN5A mRNA ARE sites in its 3′-UTR. The length of the mRNA is indicated by a kilo base (kb)-scale. (B) HuR binds to the SCN5A mRNA. After IP of RNA-protein complexes from cardiomyocytes using either anti-HuR antibody (Anti-HuR) or control IgG, RNA was isolated, and then, the cDNA was synthesized in a presence or absence of the reverse transcription enzyme. Two sets of primers to different regions of SCN5A were used to amplify SCN5A fragments. The PCR products were visualized with SYBR ™ Safe DNA Gel Stain in agarose gels. (C) qRT-PCR was performed to confirm the results in (B).

    Article Snippet: Quantitative real-time reverse-transcriptase polymerase chain reaction (qRT-PCR) was carried out using gene-specific primers, Fast SYBR® Green Master Mix and 7500 Fast Real-Time PCR System (Thermo Fisher Scientific, Waltham, MA).

    Techniques: Isolation, Synthesized, Polymerase Chain Reaction, Staining, Quantitative RT-PCR

    Performance of SYBR Green I and EvaGreen detection chemistries during qPCR assays. (A) Standard curves generated from amplification of serially diluted L . lactis phage P220 genome detected with the corresponding chemistries; (B) the corresponding performance parameters; and (C) amplification plots detected with SYBR Green I (brown) and EvaGreen (green).

    Journal: PLoS ONE

    Article Title: A high-throughput qPCR system for simultaneous quantitative detection of dairy Lactococcus lactis and Leuconostoc bacteriophages

    doi: 10.1371/journal.pone.0174223

    Figure Lengend Snippet: Performance of SYBR Green I and EvaGreen detection chemistries during qPCR assays. (A) Standard curves generated from amplification of serially diluted L . lactis phage P220 genome detected with the corresponding chemistries; (B) the corresponding performance parameters; and (C) amplification plots detected with SYBR Green I (brown) and EvaGreen (green).

    Article Snippet: Real-time qPCR assays were performed using Fast SYBR Green Master Mix (Thermo Fisher Scientific) on 7500 Fast Real-Time PCR System (Applied Biosystems, USA).

    Techniques: SYBR Green Assay, Real-time Polymerase Chain Reaction, Generated, Amplification

    Effect of variant region deletion on 3D structure and gene expression (A) Combined 3C-qPCR data of chromatin interactions in dVR cells. Black: control; Blue: dVR. The numbering of NcoI DNA fragments is given relative to the viewpoint. Viewing point: R13 region; Black bars: STARD10 exon; red bars: regulatory region; orange box: qPCR probe; orange arrow: qPCR constant primer; green stars: credible set genetic variants. Data were normalized to a CXCL12 loading control. n = 3. (B) Taqman qRT-PCR analysis of gene expression in control and dVR cells. *, P

    Journal: bioRxiv

    Article Title: Chromatin 3D interaction analysis of the STARD10 locus unveils FCHSD2 as a new regulator of insulin secretion

    doi: 10.1101/2020.03.31.017707

    Figure Lengend Snippet: Effect of variant region deletion on 3D structure and gene expression (A) Combined 3C-qPCR data of chromatin interactions in dVR cells. Black: control; Blue: dVR. The numbering of NcoI DNA fragments is given relative to the viewpoint. Viewing point: R13 region; Black bars: STARD10 exon; red bars: regulatory region; orange box: qPCR probe; orange arrow: qPCR constant primer; green stars: credible set genetic variants. Data were normalized to a CXCL12 loading control. n = 3. (B) Taqman qRT-PCR analysis of gene expression in control and dVR cells. *, P

    Article Snippet: Real-time PCR was performed on a 7500 Fast Real-Time PCR System using the Fast SYBR™ Green master mix or Fast Taqman™ master mix.

    Techniques: Variant Assay, Expressing, Real-time Polymerase Chain Reaction, Quantitative RT-PCR

    Target genes regulated by the enhancer cluster. (A) Human islet promoter capture-HiC (pcHi-C) map at STARD10 locus and surrounding region. pcHi-C interactions were considered significant with CHiCAGO > 5. Orange interactions in the pcHiC track depict interactions mediated by the CTCF sites that flank the enhancer cluster (R1 and R13). All other significant interactions are shown in dark grey. Enhancer hubs track shows all enhancers (red) and promoters (green) contained within the STARD10 hub.Orange bar in islet TADs track highlights the TAD encompassing STARD10 and surrounding genes ( ARAP1, ATG16L2 , and FCHSD2 ). (B and C) Taqman qRT-PCR analysis of gene expression in sham control and R2-deleted (dR2) cells. *, P

    Journal: bioRxiv

    Article Title: Chromatin 3D interaction analysis of the STARD10 locus unveils FCHSD2 as a new regulator of insulin secretion

    doi: 10.1101/2020.03.31.017707

    Figure Lengend Snippet: Target genes regulated by the enhancer cluster. (A) Human islet promoter capture-HiC (pcHi-C) map at STARD10 locus and surrounding region. pcHi-C interactions were considered significant with CHiCAGO > 5. Orange interactions in the pcHiC track depict interactions mediated by the CTCF sites that flank the enhancer cluster (R1 and R13). All other significant interactions are shown in dark grey. Enhancer hubs track shows all enhancers (red) and promoters (green) contained within the STARD10 hub.Orange bar in islet TADs track highlights the TAD encompassing STARD10 and surrounding genes ( ARAP1, ATG16L2 , and FCHSD2 ). (B and C) Taqman qRT-PCR analysis of gene expression in sham control and R2-deleted (dR2) cells. *, P

    Article Snippet: Real-time PCR was performed on a 7500 Fast Real-Time PCR System using the Fast SYBR™ Green master mix or Fast Taqman™ master mix.

    Techniques: Quantitative RT-PCR, Expressing

    Analysis of BMP4 and CST6 mRNA levels following treatment with demethylating agent and histone deacetylase inhibitor. (A) A CpG island (506 bp) spans the transcription start site of the human BMP4 gene. (B) Relative BMP4 mRNA levels in vehicle (DMSO)-treated 231_ATCC and 231_HM.LNm5 cells by TaqMan qRT-PCR. Expression in 231_ATCC was set to 1. Mean±s.d. ( n =3). (C) TaqMan qRT-PCR analysis of BMP4 mRNA expression levels in 5-Aza-2′-deoxycytidine (5azadC)- and Trichostatin A (TSA)-treated 231_ATCC and 231_HM.LNm5 cells. Expression in vehicle-treated cells was set to 1. Mean±s.d. ( n =3). (D) A CpG island (370 bp) spans the transcription start site of the human CST6 gene. (E) Relative CST6 mRNA levels in vehicle (DMSO)-treated 231_ATCC and 231_HM.LNm5 cells by TaqMan qRT-PCR. Expression in 231_ATCC was set to 1. Mean±s.d. ( n =3). (F) TaqMan qRT-PCR analysis of CST6 mRNA expression levels in 5azadC- and TSA-treated 231_ATCC and 231_HM.LNm5 cells. Expression in vehicle-treated cells was set to 1. Mean±s.d. ( n =3).

    Journal: Disease Models & Mechanisms

    Article Title: Functional and genomic characterisation of a xenograft model system for the study of metastasis in triple-negative breast cancer

    doi: 10.1242/dmm.032250

    Figure Lengend Snippet: Analysis of BMP4 and CST6 mRNA levels following treatment with demethylating agent and histone deacetylase inhibitor. (A) A CpG island (506 bp) spans the transcription start site of the human BMP4 gene. (B) Relative BMP4 mRNA levels in vehicle (DMSO)-treated 231_ATCC and 231_HM.LNm5 cells by TaqMan qRT-PCR. Expression in 231_ATCC was set to 1. Mean±s.d. ( n =3). (C) TaqMan qRT-PCR analysis of BMP4 mRNA expression levels in 5-Aza-2′-deoxycytidine (5azadC)- and Trichostatin A (TSA)-treated 231_ATCC and 231_HM.LNm5 cells. Expression in vehicle-treated cells was set to 1. Mean±s.d. ( n =3). (D) A CpG island (370 bp) spans the transcription start site of the human CST6 gene. (E) Relative CST6 mRNA levels in vehicle (DMSO)-treated 231_ATCC and 231_HM.LNm5 cells by TaqMan qRT-PCR. Expression in 231_ATCC was set to 1. Mean±s.d. ( n =3). (F) TaqMan qRT-PCR analysis of CST6 mRNA expression levels in 5azadC- and TSA-treated 231_ATCC and 231_HM.LNm5 cells. Expression in vehicle-treated cells was set to 1. Mean±s.d. ( n =3).

    Article Snippet: Two-step qPCR was completed (50-250 ng cDNA template/reaction) using either TaqMan gene expression assays (CTSC, ENG, TIE1, BMP2, BMP4, RPL37A) with Fast Universal PCR Master Mix, no AmpErase™ UNG (Thermo Fisher Scientific) or Fast SYBR™ Green Master Mix (50-250 ng cDNA template/reaction) and the following oligonucleotide primers (Integrated DNA Technologies, Singapore).

    Techniques: Histone Deacetylase Assay, Quantitative RT-PCR, Expressing