sybr green master mix  (Thermo Fisher)


Bioz Verified Symbol Thermo Fisher is a verified supplier  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    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
    Category:
    Kits and Assays
    Buy from Supplier


    Structured Review

    Thermo Fisher sybr green master mix
    Downregulating UPF1 from HeLa or HepG2 cells results in an upregulation of the endogenous HFE transcripts which indicates that the physiological human HFE mRNA is a natural NMD-target. HeLa cells were transiently transfected with synthetic small-interfering RNA (siRNA) duplexes directed to human UPF1 or to a non-endogenous target (Luciferase; Luc) used as control. Twenty-four (24 h), forty-eight (48 h) and seventy-two hours (72 h) after siRNA treatment, cells were harvested and protein and RNA were isolated. (A) Representative Western blot analysis of the HeLa and HepG2 cells extracts transfected with human UPF1 siRNA. Immunoblotting was performed using a human UPF1 specific antibody and a α-tubulin specific antibody to control for variations in protein loading. The percentage (%) of UPF1 protein remaining expressed in the cells after siRNA treatment is indicated below each lane and was achieved by densitometric analysis using ImageJ software. (B) Relative changes in HFE mRNA levels were analyzed by quantitative reverse transcription-coupled real-time <t>PCR</t> (RT-qPCR), normalized to the levels of endogenous G protein pathway suppressor 1 ( GPS1 ) mRNA. For that, cDNA was synthesized from total RNA and then, each cDNA sample was used as template for qPCR, which was performed using the <t>SYBR</t> Green Master Mix (Applied Biosystems) and primers that specifically hybridize to the 5′-end of HFE exon seven. Quantification of the transcript levels was performed by the absolute quantification method. Levels of HFE mRNA obtained after cellular UPF1 siRNA treatment were compared to those obtained after Luciferase siRNA treatment at the same conditions (defined as 1; arbitrary units). The histogram shows the mean and standard deviations from three independent experiments, corresponding to three independent transfections. Statistical analysis was performed using Student's t test (unpaired, two tailed). Below the histograms, there is a schematic representation of the human HFE mRNA showing its seven exons and the position of the polyadenylation [poly(A)] signals used in its 3′-end processing. The location of the initiation (AUG) and termination (STOP) codons is also represented. The double arrow represents the coordinates of the amplicon obtained in the qPCR. (C) Relative changes in HFE mRNA levels were quantified by RT-qPCR as in B but using primers that specifically hybridize to the 5′-end of the HFE exon six. Quantification of the transcript levels was performed by the absolute quantification method. Levels of HFE mRNA obtained after cellular UPF1 siRNA treatment were compared to those obtained after Luciferase siRNA treatment at the same conditions (defined as 1; arbitrary units). The histogram shows the mean and standard deviations from three independent experiments, corresponding to three independent transfections. Statistical analysis was performed as in B . Below the histograms, there is a schematic representation of the human HFE mRNA showing its seven exons and the position of the polyadenylation [poly(A)] signals used in its 3′-end processing. The location of the initiation (AUG) and termination (STOP) codons is also represented. Arrows represent the position of primers used in the qPCR. (D) Representative Western blot analysis of HeLa cells extracts transfected with human UPF1 siRNA or with a control Luciferase siRNA target. Twenty-four hours after siRNA treatment, cells were transfected with the β-globin reporter constructs (β39). Twenty-four hours post transfection, protein and RNA were isolated from the cells for analysis. Immunoblotting to confirm UPF1 knockdown was carried out with anti-UPF1 and with anti-α-tubulin antibodies (as a loading control). Identification of each band is on the right. The percentage (%) of UPF1 protein remaining expressed in the cells after siRNA treatment is indicated below each lane and was achieved as in A . (E) Relative β-globin mRNA levels in UPF1-depleted HeLa cells, normalized to the levels of puromycin resistance mRNA (plasmids carrying the reporter β-globin gene also contain the Puro r gene), were determined by quantitative RT-qPCR and compared to the corresponding β39 mRNA levels under control conditions (Luc siRNA-treated HeLa cells) (defined as 1; arbitrary units). Using the same RNA samples, relative changes in HFE mRNA levels were also quantified by RT-qPCR, using experimental conditions as in B . Levels of HFE mRNA obtained after cellular UPF1 siRNA treatment were compared to those obtained after Luciferase siRNA treatment (defined as 1; arbitrary units). The histogram shows the mean and standard deviations from three independent experiments, corresponding to three independent transfections. Statistical analysis was performed as 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/sybr green master mix/product/Thermo Fisher
    Average 99 stars, based on 5301 article reviews
    Price from $9.99 to $1999.99
    sybr green master mix - by Bioz Stars, 2020-07
    99/100 stars

    Images

    1) Product Images from "Alternative Polyadenylation and Nonsense-Mediated Decay Coordinately Regulate the Human HFE mRNA Levels"

    Article Title: Alternative Polyadenylation and Nonsense-Mediated Decay Coordinately Regulate the Human HFE mRNA Levels

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0035461

    Downregulating UPF1 from HeLa or HepG2 cells results in an upregulation of the endogenous HFE transcripts which indicates that the physiological human HFE mRNA is a natural NMD-target. HeLa cells were transiently transfected with synthetic small-interfering RNA (siRNA) duplexes directed to human UPF1 or to a non-endogenous target (Luciferase; Luc) used as control. Twenty-four (24 h), forty-eight (48 h) and seventy-two hours (72 h) after siRNA treatment, cells were harvested and protein and RNA were isolated. (A) Representative Western blot analysis of the HeLa and HepG2 cells extracts transfected with human UPF1 siRNA. Immunoblotting was performed using a human UPF1 specific antibody and a α-tubulin specific antibody to control for variations in protein loading. The percentage (%) of UPF1 protein remaining expressed in the cells after siRNA treatment is indicated below each lane and was achieved by densitometric analysis using ImageJ software. (B) Relative changes in HFE mRNA levels were analyzed by quantitative reverse transcription-coupled real-time PCR (RT-qPCR), normalized to the levels of endogenous G protein pathway suppressor 1 ( GPS1 ) mRNA. For that, cDNA was synthesized from total RNA and then, each cDNA sample was used as template for qPCR, which was performed using the SYBR Green Master Mix (Applied Biosystems) and primers that specifically hybridize to the 5′-end of HFE exon seven. Quantification of the transcript levels was performed by the absolute quantification method. Levels of HFE mRNA obtained after cellular UPF1 siRNA treatment were compared to those obtained after Luciferase siRNA treatment at the same conditions (defined as 1; arbitrary units). The histogram shows the mean and standard deviations from three independent experiments, corresponding to three independent transfections. Statistical analysis was performed using Student's t test (unpaired, two tailed). Below the histograms, there is a schematic representation of the human HFE mRNA showing its seven exons and the position of the polyadenylation [poly(A)] signals used in its 3′-end processing. The location of the initiation (AUG) and termination (STOP) codons is also represented. The double arrow represents the coordinates of the amplicon obtained in the qPCR. (C) Relative changes in HFE mRNA levels were quantified by RT-qPCR as in B but using primers that specifically hybridize to the 5′-end of the HFE exon six. Quantification of the transcript levels was performed by the absolute quantification method. Levels of HFE mRNA obtained after cellular UPF1 siRNA treatment were compared to those obtained after Luciferase siRNA treatment at the same conditions (defined as 1; arbitrary units). The histogram shows the mean and standard deviations from three independent experiments, corresponding to three independent transfections. Statistical analysis was performed as in B . Below the histograms, there is a schematic representation of the human HFE mRNA showing its seven exons and the position of the polyadenylation [poly(A)] signals used in its 3′-end processing. The location of the initiation (AUG) and termination (STOP) codons is also represented. Arrows represent the position of primers used in the qPCR. (D) Representative Western blot analysis of HeLa cells extracts transfected with human UPF1 siRNA or with a control Luciferase siRNA target. Twenty-four hours after siRNA treatment, cells were transfected with the β-globin reporter constructs (β39). Twenty-four hours post transfection, protein and RNA were isolated from the cells for analysis. Immunoblotting to confirm UPF1 knockdown was carried out with anti-UPF1 and with anti-α-tubulin antibodies (as a loading control). Identification of each band is on the right. The percentage (%) of UPF1 protein remaining expressed in the cells after siRNA treatment is indicated below each lane and was achieved as in A . (E) Relative β-globin mRNA levels in UPF1-depleted HeLa cells, normalized to the levels of puromycin resistance mRNA (plasmids carrying the reporter β-globin gene also contain the Puro r gene), were determined by quantitative RT-qPCR and compared to the corresponding β39 mRNA levels under control conditions (Luc siRNA-treated HeLa cells) (defined as 1; arbitrary units). Using the same RNA samples, relative changes in HFE mRNA levels were also quantified by RT-qPCR, using experimental conditions as in B . Levels of HFE mRNA obtained after cellular UPF1 siRNA treatment were compared to those obtained after Luciferase siRNA treatment (defined as 1; arbitrary units). The histogram shows the mean and standard deviations from three independent experiments, corresponding to three independent transfections. Statistical analysis was performed as in B .
    Figure Legend Snippet: Downregulating UPF1 from HeLa or HepG2 cells results in an upregulation of the endogenous HFE transcripts which indicates that the physiological human HFE mRNA is a natural NMD-target. HeLa cells were transiently transfected with synthetic small-interfering RNA (siRNA) duplexes directed to human UPF1 or to a non-endogenous target (Luciferase; Luc) used as control. Twenty-four (24 h), forty-eight (48 h) and seventy-two hours (72 h) after siRNA treatment, cells were harvested and protein and RNA were isolated. (A) Representative Western blot analysis of the HeLa and HepG2 cells extracts transfected with human UPF1 siRNA. Immunoblotting was performed using a human UPF1 specific antibody and a α-tubulin specific antibody to control for variations in protein loading. The percentage (%) of UPF1 protein remaining expressed in the cells after siRNA treatment is indicated below each lane and was achieved by densitometric analysis using ImageJ software. (B) Relative changes in HFE mRNA levels were analyzed by quantitative reverse transcription-coupled real-time PCR (RT-qPCR), normalized to the levels of endogenous G protein pathway suppressor 1 ( GPS1 ) mRNA. For that, cDNA was synthesized from total RNA and then, each cDNA sample was used as template for qPCR, which was performed using the SYBR Green Master Mix (Applied Biosystems) and primers that specifically hybridize to the 5′-end of HFE exon seven. Quantification of the transcript levels was performed by the absolute quantification method. Levels of HFE mRNA obtained after cellular UPF1 siRNA treatment were compared to those obtained after Luciferase siRNA treatment at the same conditions (defined as 1; arbitrary units). The histogram shows the mean and standard deviations from three independent experiments, corresponding to three independent transfections. Statistical analysis was performed using Student's t test (unpaired, two tailed). Below the histograms, there is a schematic representation of the human HFE mRNA showing its seven exons and the position of the polyadenylation [poly(A)] signals used in its 3′-end processing. The location of the initiation (AUG) and termination (STOP) codons is also represented. The double arrow represents the coordinates of the amplicon obtained in the qPCR. (C) Relative changes in HFE mRNA levels were quantified by RT-qPCR as in B but using primers that specifically hybridize to the 5′-end of the HFE exon six. Quantification of the transcript levels was performed by the absolute quantification method. Levels of HFE mRNA obtained after cellular UPF1 siRNA treatment were compared to those obtained after Luciferase siRNA treatment at the same conditions (defined as 1; arbitrary units). The histogram shows the mean and standard deviations from three independent experiments, corresponding to three independent transfections. Statistical analysis was performed as in B . Below the histograms, there is a schematic representation of the human HFE mRNA showing its seven exons and the position of the polyadenylation [poly(A)] signals used in its 3′-end processing. The location of the initiation (AUG) and termination (STOP) codons is also represented. Arrows represent the position of primers used in the qPCR. (D) Representative Western blot analysis of HeLa cells extracts transfected with human UPF1 siRNA or with a control Luciferase siRNA target. Twenty-four hours after siRNA treatment, cells were transfected with the β-globin reporter constructs (β39). Twenty-four hours post transfection, protein and RNA were isolated from the cells for analysis. Immunoblotting to confirm UPF1 knockdown was carried out with anti-UPF1 and with anti-α-tubulin antibodies (as a loading control). Identification of each band is on the right. The percentage (%) of UPF1 protein remaining expressed in the cells after siRNA treatment is indicated below each lane and was achieved as in A . (E) Relative β-globin mRNA levels in UPF1-depleted HeLa cells, normalized to the levels of puromycin resistance mRNA (plasmids carrying the reporter β-globin gene also contain the Puro r gene), were determined by quantitative RT-qPCR and compared to the corresponding β39 mRNA levels under control conditions (Luc siRNA-treated HeLa cells) (defined as 1; arbitrary units). Using the same RNA samples, relative changes in HFE mRNA levels were also quantified by RT-qPCR, using experimental conditions as in B . Levels of HFE mRNA obtained after cellular UPF1 siRNA treatment were compared to those obtained after Luciferase siRNA treatment (defined as 1; arbitrary units). The histogram shows the mean and standard deviations from three independent experiments, corresponding to three independent transfections. Statistical analysis was performed as in B .

    Techniques Used: Transfection, Small Interfering RNA, Luciferase, Isolation, Western Blot, Software, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Synthesized, SYBR Green Assay, Two Tailed Test, Amplification, Construct

    2) Product Images from "Ovarian carcinomas with genetic and epigenetic BRCA1 loss have distinct molecular abnormalities"

    Article Title: Ovarian carcinomas with genetic and epigenetic BRCA1 loss have distinct molecular abnormalities

    Journal: BMC Cancer

    doi: 10.1186/1471-2407-8-17

    Assessment of BRCA1 loss (A) Mutation screening showing the abnormal denaturing high performance liquid chromatography profile corresponding to the 1351delAT mutation in tumor 223. The single blue line represents the electropherogram from a normal control, while the purple line represents the abnormal profile formed by the mutated exon 11c in tumor 223. (B) Direct DNA sequencing demonstrating the 185delAG mutation in tumor 283. Only the mutant allele is seen in the tumor because LOH is present. (C-E) Loss of heterozygosity (LOH) analysis using BRCA1-associated microsatellite markers visualized on an ABI Prism 3100 Genetic Analyzer, where LOH is defined as > 50% decrease in area under the curve when germline DNA (upper tracing) and tumor DNA (lower tracing) are compared. (C) The lack of LOH in tumor 240 demonstrated using microsatellite marker D17S1185, (D) LOH in tumor 283 demonstrated using microsatellite marker D17S855. (E) Microsatellite instability demonstrated in tumor 156 using microsatellite marker D17S1185. (F, G, H, and I) Methylation analysis of BRCA1 gene using fluorescence-based, quantitative, real-time PCR (TaqMan) using SYBR Green 1 as detection method. Two sets of primers, designed specifically for bisulfite converted DNA, were used: a methylated set for the BRCA1 gene and a reference set (MYOD1) to control for input DNA. Specificity of the reactions for methylated DNA were confirmed separately using human genomic DNA (unmethyated; F) and CpG methylated Jurkat genomic DNA (methylated; G), respectively. H and I show representative examples of results from assessment of BRCA1 loss through promoter hypermethyation. Tumor 178 shows only unmethylated BRCA1 promoter, while tumor 345 shows evidence of BRCA1 promoter hypermethylation.
    Figure Legend Snippet: Assessment of BRCA1 loss (A) Mutation screening showing the abnormal denaturing high performance liquid chromatography profile corresponding to the 1351delAT mutation in tumor 223. The single blue line represents the electropherogram from a normal control, while the purple line represents the abnormal profile formed by the mutated exon 11c in tumor 223. (B) Direct DNA sequencing demonstrating the 185delAG mutation in tumor 283. Only the mutant allele is seen in the tumor because LOH is present. (C-E) Loss of heterozygosity (LOH) analysis using BRCA1-associated microsatellite markers visualized on an ABI Prism 3100 Genetic Analyzer, where LOH is defined as > 50% decrease in area under the curve when germline DNA (upper tracing) and tumor DNA (lower tracing) are compared. (C) The lack of LOH in tumor 240 demonstrated using microsatellite marker D17S1185, (D) LOH in tumor 283 demonstrated using microsatellite marker D17S855. (E) Microsatellite instability demonstrated in tumor 156 using microsatellite marker D17S1185. (F, G, H, and I) Methylation analysis of BRCA1 gene using fluorescence-based, quantitative, real-time PCR (TaqMan) using SYBR Green 1 as detection method. Two sets of primers, designed specifically for bisulfite converted DNA, were used: a methylated set for the BRCA1 gene and a reference set (MYOD1) to control for input DNA. Specificity of the reactions for methylated DNA were confirmed separately using human genomic DNA (unmethyated; F) and CpG methylated Jurkat genomic DNA (methylated; G), respectively. H and I show representative examples of results from assessment of BRCA1 loss through promoter hypermethyation. Tumor 178 shows only unmethylated BRCA1 promoter, while tumor 345 shows evidence of BRCA1 promoter hypermethylation.

    Techniques Used: Mutagenesis, High Performance Liquid Chromatography, DNA Sequencing, Marker, Methylation, Fluorescence, Real-time Polymerase Chain Reaction, SYBR Green Assay

    3) Product Images from "NFAT2 Regulates Generation of Innate-Like CD8+ T Lymphocytes and CD8+ T Lymphocytes Responses"

    Article Title: NFAT2 Regulates Generation of Innate-Like CD8+ T Lymphocytes and CD8+ T Lymphocytes Responses

    Journal: Frontiers in Immunology

    doi: 10.3389/fimmu.2016.00411

    IFN-γ production by cytotoxic CD8 + T cells is regulated by NFAT2 . (A) Schematic representation of cytotoxic CD8 + T cell differentiation in vitro . Purified CD8 + T cells from WT and NFAT2 fl/fl CD4-Cre mice were differentiated in vitro into cytotoxic CD8 + T lymphocytes as described. (B) Total RNA isolated from naive CD8 + T cells (NAIVE) or from in vitro differentiated cytotoxic CD8+ T lymphocytes (CTLs) were analyzed for NFAT2 mRNA levels by real-time RT-PCR assay using SYBR green master mix. The data are normalized to the β-actin levels. (C) Total number of live WT and NFAT2-deficient CD8 + T lymphocytes was recorded daily during in vitro differentiation using Trypan blue exclusion method. (D) Flow cytometric analysis of CD44 and CD25 expression on differentiated cytotoxic WT and NFAT2-deficient CD8 + T cells at day 5. The analysis of MFI from three independent experiments on the right. (E) At day 5, differentiated cytotoxic CD8 + T cells were tested in cytotoxicity assay against P815 cells as described at indicated efector:target ratios or restimulated with PMA plus ionomycin for 6 h, and intracellular granzime B and IFN-γ production was analyzed by flow cytometry. The analysis of MFI from three independent experiments on the right (F) . (G) Total RNA was isolated from differentiated in vitro WT and NFAT2-deficient cytotoxic CD8 + T lymphocytes, and T-bet and Blimp-1 mRNA levels were analyzed by real-time RT-PCR assay using Taqman probes. The data are normalized to the HPRT RNA levels. All data are shown as mean ± SD of three independent experiments. The ns indicates not significant and *indicates p
    Figure Legend Snippet: IFN-γ production by cytotoxic CD8 + T cells is regulated by NFAT2 . (A) Schematic representation of cytotoxic CD8 + T cell differentiation in vitro . Purified CD8 + T cells from WT and NFAT2 fl/fl CD4-Cre mice were differentiated in vitro into cytotoxic CD8 + T lymphocytes as described. (B) Total RNA isolated from naive CD8 + T cells (NAIVE) or from in vitro differentiated cytotoxic CD8+ T lymphocytes (CTLs) were analyzed for NFAT2 mRNA levels by real-time RT-PCR assay using SYBR green master mix. The data are normalized to the β-actin levels. (C) Total number of live WT and NFAT2-deficient CD8 + T lymphocytes was recorded daily during in vitro differentiation using Trypan blue exclusion method. (D) Flow cytometric analysis of CD44 and CD25 expression on differentiated cytotoxic WT and NFAT2-deficient CD8 + T cells at day 5. The analysis of MFI from three independent experiments on the right. (E) At day 5, differentiated cytotoxic CD8 + T cells were tested in cytotoxicity assay against P815 cells as described at indicated efector:target ratios or restimulated with PMA plus ionomycin for 6 h, and intracellular granzime B and IFN-γ production was analyzed by flow cytometry. The analysis of MFI from three independent experiments on the right (F) . (G) Total RNA was isolated from differentiated in vitro WT and NFAT2-deficient cytotoxic CD8 + T lymphocytes, and T-bet and Blimp-1 mRNA levels were analyzed by real-time RT-PCR assay using Taqman probes. The data are normalized to the HPRT RNA levels. All data are shown as mean ± SD of three independent experiments. The ns indicates not significant and *indicates p

    Techniques Used: Cell Differentiation, In Vitro, Purification, Mouse Assay, Isolation, Quantitative RT-PCR, SYBR Green Assay, Flow Cytometry, Expressing, Cytotoxicity Assay, Cytometry

    NFAT2 is present and functional in CD8 + T lymphocytes . CD8 + T lymphocytes were purified from naive C57BL/6 mice, as described, and then left unstimulated (0) or stimulated in vitro with anti-CD3 plus anti-CD28 (both at 1 μg/ml) for indicated times. (A) Total RNA was isolated, and NFAT2 mRNA levels were analyzed by real-time RT-PCR assay using SYBR green master mix. The data are normalized to the β-actin levels. Data are shown as mean ± SD of three independent experiments. Detection of NFAT2 transcription factor in CD8 + T cells in total lysates (B) and in cytoplasmic and nuclear fractions (C) by Western blot. (D) Cellular localization of NFAT2 protein in CD8 + T cells by immunofluorescence staining. All data are representative of at least two independent experiments.
    Figure Legend Snippet: NFAT2 is present and functional in CD8 + T lymphocytes . CD8 + T lymphocytes were purified from naive C57BL/6 mice, as described, and then left unstimulated (0) or stimulated in vitro with anti-CD3 plus anti-CD28 (both at 1 μg/ml) for indicated times. (A) Total RNA was isolated, and NFAT2 mRNA levels were analyzed by real-time RT-PCR assay using SYBR green master mix. The data are normalized to the β-actin levels. Data are shown as mean ± SD of three independent experiments. Detection of NFAT2 transcription factor in CD8 + T cells in total lysates (B) and in cytoplasmic and nuclear fractions (C) by Western blot. (D) Cellular localization of NFAT2 protein in CD8 + T cells by immunofluorescence staining. All data are representative of at least two independent experiments.

    Techniques Used: Functional Assay, Purification, Mouse Assay, In Vitro, Isolation, Quantitative RT-PCR, SYBR Green Assay, Western Blot, Immunofluorescence, Staining

    4) Product Images from "p21Waf1 expression is regulated by nuclear intermediate filament vimentin in neuroblastoma"

    Article Title: p21Waf1 expression is regulated by nuclear intermediate filament vimentin in neuroblastoma

    Journal: BMC Cancer

    doi: 10.1186/1471-2407-10-473

    Vimentin-p21 mRNA correlation in NB cell lines and tumors - Association between p21 expression and clinical evolution . A. mRNA quantification of vimentin ( left panel ) and p21 ( right panel ) relative expression level after GAPDH normalization, in 10 high risk NB cell lines, using Sybr Green Q-PCR. The S- or N-subtype of the NB cell lines is indicated. This result was confirmed after EF1α normalization and by TaqMan Q-PCR quantification, using two different genes for normalization (18 S and TFRC) (data not shown). Bars and error bars represent the means and standard deviations, respectively. B. Representation of the p21 mRNA relative expression level according to that of vimentin, quantified by Sybr Green Q-PCR after 18 S normalization, in 77 NB tumor samples collected at the time of diagnosis (primary tumor biopsies or metastatic bone marrows, all containing more than 70% of tumor cells) (Clinical data are presented in Additional file 1 , Table S1). A cDNA sample from the SK-N-SH cell line was used as a reference to integrate all the data together in a unique analysis. The Spearman statistical test was performed with the Graph Pad Prism Software version 4.0. CI: Confidence Interval. C. Left panel: Representation of the p21 mRNA relative expression level according to that of vimentin, quantified by TaqMan Q-PCR after 18 S normalization, in the 40 high risk NB tumor samples of the cohort. The Spearman statistical test was performed as in 1B. This result was obtained using either Sybr-Green or TaqMan technology with two normalization genes, 18 S and GAPDH, or 18 S and TFRC, respectively (data not shown). Right panel: The survival analysis of patients older than one year of age with a stage 4 NB (N = 40) was performed using the Log-rank Kaplan-Meier estimation with the Graph Pad Prism Software version 4.0. The overall survival was defined as the time in months between diagnosis and death or last follow-up (excluding patients who died from toxicity). Patients were separated into two distinct groups according to their p21 mRNA tumor relative expression level quantified by Q-PCR as in the left panel. The median of p21 tumor expression was chosen as a cut-off between high ( > median, black line) and low (
    Figure Legend Snippet: Vimentin-p21 mRNA correlation in NB cell lines and tumors - Association between p21 expression and clinical evolution . A. mRNA quantification of vimentin ( left panel ) and p21 ( right panel ) relative expression level after GAPDH normalization, in 10 high risk NB cell lines, using Sybr Green Q-PCR. The S- or N-subtype of the NB cell lines is indicated. This result was confirmed after EF1α normalization and by TaqMan Q-PCR quantification, using two different genes for normalization (18 S and TFRC) (data not shown). Bars and error bars represent the means and standard deviations, respectively. B. Representation of the p21 mRNA relative expression level according to that of vimentin, quantified by Sybr Green Q-PCR after 18 S normalization, in 77 NB tumor samples collected at the time of diagnosis (primary tumor biopsies or metastatic bone marrows, all containing more than 70% of tumor cells) (Clinical data are presented in Additional file 1 , Table S1). A cDNA sample from the SK-N-SH cell line was used as a reference to integrate all the data together in a unique analysis. The Spearman statistical test was performed with the Graph Pad Prism Software version 4.0. CI: Confidence Interval. C. Left panel: Representation of the p21 mRNA relative expression level according to that of vimentin, quantified by TaqMan Q-PCR after 18 S normalization, in the 40 high risk NB tumor samples of the cohort. The Spearman statistical test was performed as in 1B. This result was obtained using either Sybr-Green or TaqMan technology with two normalization genes, 18 S and GAPDH, or 18 S and TFRC, respectively (data not shown). Right panel: The survival analysis of patients older than one year of age with a stage 4 NB (N = 40) was performed using the Log-rank Kaplan-Meier estimation with the Graph Pad Prism Software version 4.0. The overall survival was defined as the time in months between diagnosis and death or last follow-up (excluding patients who died from toxicity). Patients were separated into two distinct groups according to their p21 mRNA tumor relative expression level quantified by Q-PCR as in the left panel. The median of p21 tumor expression was chosen as a cut-off between high ( > median, black line) and low (

    Techniques Used: Expressing, SYBR Green Assay, Polymerase Chain Reaction, Software

    5) Product Images from "The Lipopolysaccharide and ?-1,3-Glucan Binding Protein Gene Is Upregulated in White Spot Virus-Infected Shrimp (Penaeus stylirostris)"

    Article Title: The Lipopolysaccharide and ?-1,3-Glucan Binding Protein Gene Is Upregulated in White Spot Virus-Infected Shrimp (Penaeus stylirostris)

    Journal: Journal of Virology

    doi: 10.1128/JVI.76.14.7140-7149.2002

    Temporal expression of the LGBP gene in the hepatopancreas of healthy and WSV-injected P. stylirostris as measured by real-time quantitative RT-PCR. The mRNA expression of the LGBP gene and the internal control EF-1α gene was measured by SYBR Green RT-PCR in each healthy ( n = 7) and WSV-injected ( n = 7) shrimp at 0, 2, 4, 8, 16, and 32 h p.i. The relative LGBP level as expressed by Δ C T ( C T value of LGBP gene − C T value of EF-1α gene) was determined for each sample. The average Δ C T for healthy (⋄) and WSV-injected (▪) samples for each time point was then used to plot the graph.
    Figure Legend Snippet: Temporal expression of the LGBP gene in the hepatopancreas of healthy and WSV-injected P. stylirostris as measured by real-time quantitative RT-PCR. The mRNA expression of the LGBP gene and the internal control EF-1α gene was measured by SYBR Green RT-PCR in each healthy ( n = 7) and WSV-injected ( n = 7) shrimp at 0, 2, 4, 8, 16, and 32 h p.i. The relative LGBP level as expressed by Δ C T ( C T value of LGBP gene − C T value of EF-1α gene) was determined for each sample. The average Δ C T for healthy (⋄) and WSV-injected (▪) samples for each time point was then used to plot the graph.

    Techniques Used: Expressing, Injection, Quantitative RT-PCR, SYBR Green Assay, Reverse Transcription Polymerase Chain Reaction

    Measurement of LGBP mRNA expression in healthy and WSV-injected P. stylirostris shrimp by SYBR Green RT-PCR in two independent WSV challenge experiments. Each bar represents the average relative level of LGBP mRNA as measured by SYBR Green RT-PCR. The expression of LGBP and EF-1α genes was measured in healthy and WSV-injected shrimp at 32 h p.i. The Δ C T ( C T value of LGBP gene − C T value of EF-1α gene) was measured for each healthy ( n = 7) and WSV-infected ( n = 7) animal, and the average Δ C T values were used to draw the bars. The numbers above the bars indicate the average Δ C T values. A 3.3-Δ C T -value difference (ΔΔ C T ) indicates a 10-fold difference between the two treatments. C T values and the copy number of the genes are inversely related, and therefore, a sample with higher C T values contains less of the target gene.
    Figure Legend Snippet: Measurement of LGBP mRNA expression in healthy and WSV-injected P. stylirostris shrimp by SYBR Green RT-PCR in two independent WSV challenge experiments. Each bar represents the average relative level of LGBP mRNA as measured by SYBR Green RT-PCR. The expression of LGBP and EF-1α genes was measured in healthy and WSV-injected shrimp at 32 h p.i. The Δ C T ( C T value of LGBP gene − C T value of EF-1α gene) was measured for each healthy ( n = 7) and WSV-infected ( n = 7) animal, and the average Δ C T values were used to draw the bars. The numbers above the bars indicate the average Δ C T values. A 3.3-Δ C T -value difference (ΔΔ C T ) indicates a 10-fold difference between the two treatments. C T values and the copy number of the genes are inversely related, and therefore, a sample with higher C T values contains less of the target gene.

    Techniques Used: Expressing, Injection, SYBR Green Assay, Reverse Transcription Polymerase Chain Reaction, Infection

    Amplification profiles (A and C) and dissociation curves (B and D) of the LGBP gene (A and B) and the internal control EF-1α gene (C and D). The mRNA expression of LGBP and EF-1α genes in healthy and WSV-infected shrimp at 32 h p.i. was quantified by SYBR Green RT-PCR. The melting temperatures ( T m ) of the amplicons are indicated alongside their dissociation curves.
    Figure Legend Snippet: Amplification profiles (A and C) and dissociation curves (B and D) of the LGBP gene (A and B) and the internal control EF-1α gene (C and D). The mRNA expression of LGBP and EF-1α genes in healthy and WSV-infected shrimp at 32 h p.i. was quantified by SYBR Green RT-PCR. The melting temperatures ( T m ) of the amplicons are indicated alongside their dissociation curves.

    Techniques Used: Amplification, Expressing, Infection, SYBR Green Assay, Reverse Transcription Polymerase Chain Reaction

    6) Product Images from "Junction region of EWS-FLI1 fusion protein has a dominant negative effect in Ewing's Sarcoma in vitro"

    Article Title: Junction region of EWS-FLI1 fusion protein has a dominant negative effect in Ewing's Sarcoma in vitro

    Journal: BMC Cancer

    doi: 10.1186/1471-2407-12-513

    Over-expression of EWS-FLI1 junction region (aa 251- aa 280) inhibits tumorigenicity, EWS-FLI1 target gene expression and EMT marker genes in Ewing’s sarcoma cells. A : Confirmation of junction region (aa 251–280) protein expression in transfected EWS502 cell line by immunoblotting with anti Flag antibody. Lane 1: pCDNA/FLAG expressing EWS502 cells, Lane 2: pCDNA/FLAG/Junction (aa 251–280) expressing EWS502 cells. Figure 5 B : Real time RT-PCR analysis of Junction region (aa 251–280) in pCDNA/FLAG and pCDNA/FLAG/Junction (aa 251–280) stably expressing EWS502 cells. The real time RT-PCR analysis was performed using the same primer set used to clone the junction region and SYBR green chemistry. Figure 5 C : Soft agar assay showing significant reduction in colony number in EWS502 transfected with junction deletion construct, compared to vector control and wild type EWS502. Figure 5 D : Photographs of live pCDNA/FLAG and pCDNA/FLAG/Junction (aa 251–280) stably expressing EWS502 cells in culture obtained at 10X magnification.
    Figure Legend Snippet: Over-expression of EWS-FLI1 junction region (aa 251- aa 280) inhibits tumorigenicity, EWS-FLI1 target gene expression and EMT marker genes in Ewing’s sarcoma cells. A : Confirmation of junction region (aa 251–280) protein expression in transfected EWS502 cell line by immunoblotting with anti Flag antibody. Lane 1: pCDNA/FLAG expressing EWS502 cells, Lane 2: pCDNA/FLAG/Junction (aa 251–280) expressing EWS502 cells. Figure 5 B : Real time RT-PCR analysis of Junction region (aa 251–280) in pCDNA/FLAG and pCDNA/FLAG/Junction (aa 251–280) stably expressing EWS502 cells. The real time RT-PCR analysis was performed using the same primer set used to clone the junction region and SYBR green chemistry. Figure 5 C : Soft agar assay showing significant reduction in colony number in EWS502 transfected with junction deletion construct, compared to vector control and wild type EWS502. Figure 5 D : Photographs of live pCDNA/FLAG and pCDNA/FLAG/Junction (aa 251–280) stably expressing EWS502 cells in culture obtained at 10X magnification.

    Techniques Used: Over Expression, Expressing, Marker, Transfection, Quantitative RT-PCR, Stable Transfection, SYBR Green Assay, Soft Agar Assay, Construct, Plasmid Preparation

    7) Product Images from "The effects of black garlic (Allium satvium) extracts on lipid metabolism in rats fed a high fat diet"

    Article Title: The effects of black garlic (Allium satvium) extracts on lipid metabolism in rats fed a high fat diet

    Journal: Nutrition Research and Practice

    doi: 10.4162/nrp.2015.9.1.30

    Effect of black garlic extracts on mRNA expression of protein related to cholesterol synthesis in liver of rats. Total RNA was isolated using TRI-reagenet and cDNA was synthesized using 3 µg of total RNA with SuperScript II reverse transcriptase. Realtime PCR was performed using SYBR green and standard procedures to assess the mRNA expression of primer in liver samples obtained from each group. An Applied Biosystem StepOne softwere v2.1 was used. Each bar represents the mean ± SE of three independent experiments. Different letters above each bar indicate significant differences among groups at α = 0.05 as determined by Duncan's multiple range tests. ACAT; Acyl-CoA cholesterol acyltransferase, HMG-CoA; Hydroxy-3-methylglutaryl coenzyme A
    Figure Legend Snippet: Effect of black garlic extracts on mRNA expression of protein related to cholesterol synthesis in liver of rats. Total RNA was isolated using TRI-reagenet and cDNA was synthesized using 3 µg of total RNA with SuperScript II reverse transcriptase. Realtime PCR was performed using SYBR green and standard procedures to assess the mRNA expression of primer in liver samples obtained from each group. An Applied Biosystem StepOne softwere v2.1 was used. Each bar represents the mean ± SE of three independent experiments. Different letters above each bar indicate significant differences among groups at α = 0.05 as determined by Duncan's multiple range tests. ACAT; Acyl-CoA cholesterol acyltransferase, HMG-CoA; Hydroxy-3-methylglutaryl coenzyme A

    Techniques Used: Expressing, Isolation, Synthesized, Polymerase Chain Reaction, SYBR Green Assay

    Effect of black garlic extracts on mRNA expression of CPT1 in liver of rats. Total RNA was isolated using TRI-reagenet and cDNA was synthesized using 3 µg of total RNA with SuperScript II reverse transcriptase. Realtime PCR was performed using SYBR green and standard procedures to assess the mRNA expression of primer in liver samples obtained from each group. An Applied Biosystem StepOne softwere v2.1 was used. Each bar represents the mean ± SE of three independent experiments. Different letters above each bar indicate significant differences among groups at α = 0.05 as determined by Duncan's multiple range test. CPT1; Carnitine palmitoyltransferase-1
    Figure Legend Snippet: Effect of black garlic extracts on mRNA expression of CPT1 in liver of rats. Total RNA was isolated using TRI-reagenet and cDNA was synthesized using 3 µg of total RNA with SuperScript II reverse transcriptase. Realtime PCR was performed using SYBR green and standard procedures to assess the mRNA expression of primer in liver samples obtained from each group. An Applied Biosystem StepOne softwere v2.1 was used. Each bar represents the mean ± SE of three independent experiments. Different letters above each bar indicate significant differences among groups at α = 0.05 as determined by Duncan's multiple range test. CPT1; Carnitine palmitoyltransferase-1

    Techniques Used: Expressing, Isolation, Synthesized, Polymerase Chain Reaction, SYBR Green Assay

    Effect of black garlic extracts on mRNA expression of transcription factor and enzymes related to fat synthesis in liver of rats. Total RNA was isolated using TRI-reagenet and cDNA was synthesized using 3 µg of total RNA with SuperScript II reverse transcriptase. Realtime PCR was performed using SYBR green and standard procedures to assess the mRNA expression of primer in liver samples obtained from each group. An Applied Biosystem StepOne softwere v2.1 was used. Each bar represents the mean ± SE of three independent experiments. Different letters above each bar indicate significant differences among groups at α = 0.05 as determined by Duncan's multiple range test. ACC; Acetyl-CoA carboxylase, FAS; Fatty acid synthase, G6PDH; Glucose-6-phosphate dehydrogenase, SREBP-1C; Sterol regulatory element binding protein
    Figure Legend Snippet: Effect of black garlic extracts on mRNA expression of transcription factor and enzymes related to fat synthesis in liver of rats. Total RNA was isolated using TRI-reagenet and cDNA was synthesized using 3 µg of total RNA with SuperScript II reverse transcriptase. Realtime PCR was performed using SYBR green and standard procedures to assess the mRNA expression of primer in liver samples obtained from each group. An Applied Biosystem StepOne softwere v2.1 was used. Each bar represents the mean ± SE of three independent experiments. Different letters above each bar indicate significant differences among groups at α = 0.05 as determined by Duncan's multiple range test. ACC; Acetyl-CoA carboxylase, FAS; Fatty acid synthase, G6PDH; Glucose-6-phosphate dehydrogenase, SREBP-1C; Sterol regulatory element binding protein

    Techniques Used: Expressing, Isolation, Synthesized, Polymerase Chain Reaction, SYBR Green Assay, Binding Assay

    8) Product Images from "Methamphetamine mediates immune dysregulation in a murine model of chronic viral infection"

    Article Title: Methamphetamine mediates immune dysregulation in a murine model of chronic viral infection

    Journal: Frontiers in Microbiology

    doi: 10.3389/fmicb.2015.00793

    METH treatment does not alter plasma or organ viral load in chronic LCMV infection . LCMV viral load in plasma (A) , spleen (B) , and lungs (C) was measured by RT-PCR. Real-time quantitative PCR of LCMV viral cDNA was performed using LCMV GP primers and Sybr Green Master Mix. A standard curve was generated from the LCMV-Arm53b plasmid to calculate the copy numbers. Samples and standards were performed in triplicate with five animals per experimental condition.
    Figure Legend Snippet: METH treatment does not alter plasma or organ viral load in chronic LCMV infection . LCMV viral load in plasma (A) , spleen (B) , and lungs (C) was measured by RT-PCR. Real-time quantitative PCR of LCMV viral cDNA was performed using LCMV GP primers and Sybr Green Master Mix. A standard curve was generated from the LCMV-Arm53b plasmid to calculate the copy numbers. Samples and standards were performed in triplicate with five animals per experimental condition.

    Techniques Used: Infection, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction, SYBR Green Assay, Generated, Plasmid Preparation

    9) Product Images from "Deciduous DPSCs Ameliorate MPTP-Mediated Neurotoxicity, Sensorimotor Coordination and Olfactory Function in Parkinsonian Mice"

    Article Title: Deciduous DPSCs Ameliorate MPTP-Mediated Neurotoxicity, Sensorimotor Coordination and Olfactory Function in Parkinsonian Mice

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms20030568

    Detection of pluripotent indicators as well as neuronal markers. The Ct value of genes was analysed in the study using SYBR green-based qRT–PCR for DPSCs. Generally, the higher a fold change value, the more copies are present in the specific sample. Total RNA from brain was used as a positive control. Values are presented after normalization to 18s mRNA levels ( p
    Figure Legend Snippet: Detection of pluripotent indicators as well as neuronal markers. The Ct value of genes was analysed in the study using SYBR green-based qRT–PCR for DPSCs. Generally, the higher a fold change value, the more copies are present in the specific sample. Total RNA from brain was used as a positive control. Values are presented after normalization to 18s mRNA levels ( p

    Techniques Used: SYBR Green Assay, Quantitative RT-PCR, Positive Control

    10) Product Images from "The Genetic and Molecular Basis of O-Antigenic Diversity in Burkholderia pseudomallei Lipopolysaccharide"

    Article Title: The Genetic and Molecular Basis of O-Antigenic Diversity in Burkholderia pseudomallei Lipopolysaccharide

    Journal: PLoS Neglected Tropical Diseases

    doi: 10.1371/journal.pntd.0001453

    Genotyping scheme and frequencies of three different LPS genotypes identified in B. pseudomallei populations. Multiplex SYBR-Green PCR assays were developed to target the presence of genes: wbiE , BUC_3396, and BURP840_LPSb16, which were the representatives of LPS genotypes A, B, and B2, respectively. PCR amplicons from these 3 gene targets were differentiated by melting dissociation (A); or sizing (B); lanes 1, 2, 3, and 4 are PCR products from strains K96243, 576, MSHR840, and non-DNA template control (NTC), respectively; and L, 1 kb-plus DNA ladder. We note that LPS genotype A was the most common LPS genotype, whereas a majority of the LPS genotype B was found in strains from Australia (approx.13.8%). Genotype B2 was found in strains from Australia and Papua New Guinea (PNG) only (C).
    Figure Legend Snippet: Genotyping scheme and frequencies of three different LPS genotypes identified in B. pseudomallei populations. Multiplex SYBR-Green PCR assays were developed to target the presence of genes: wbiE , BUC_3396, and BURP840_LPSb16, which were the representatives of LPS genotypes A, B, and B2, respectively. PCR amplicons from these 3 gene targets were differentiated by melting dissociation (A); or sizing (B); lanes 1, 2, 3, and 4 are PCR products from strains K96243, 576, MSHR840, and non-DNA template control (NTC), respectively; and L, 1 kb-plus DNA ladder. We note that LPS genotype A was the most common LPS genotype, whereas a majority of the LPS genotype B was found in strains from Australia (approx.13.8%). Genotype B2 was found in strains from Australia and Papua New Guinea (PNG) only (C).

    Techniques Used: Multiplex Assay, SYBR Green Assay, Polymerase Chain Reaction

    11) Product Images from "GAPDH, β-actin and β2-microglobulin, as three common reference genes, are not reliable for gene expression studies in equine adipose- and marrow-derived mesenchymal stem cells"

    Article Title: GAPDH, β-actin and β2-microglobulin, as three common reference genes, are not reliable for gene expression studies in equine adipose- and marrow-derived mesenchymal stem cells

    Journal: Journal of Animal Science and Technology

    doi: 10.1186/s40781-015-0050-8

    Derivative melting-curve analysis using SYBR Green to identify the dissociation temperature of reference gene amplicons and the specificity of RT-PCR reactions using total RNA from equine marrow- and adipose derived MSCs. The Optical System Software plotted the rate of change of the relative fluorescence units with time (dF)/dT) on the Y-axis versus the temperature (°C) on the X-axis, which peaks at the melting temperature (Tm). Similar peaks indicate no contamination and primer–dimer artifact. BM = Bone marrow; AT = Adipose tissue; MSCs = Mesenchymal stem cells.
    Figure Legend Snippet: Derivative melting-curve analysis using SYBR Green to identify the dissociation temperature of reference gene amplicons and the specificity of RT-PCR reactions using total RNA from equine marrow- and adipose derived MSCs. The Optical System Software plotted the rate of change of the relative fluorescence units with time (dF)/dT) on the Y-axis versus the temperature (°C) on the X-axis, which peaks at the melting temperature (Tm). Similar peaks indicate no contamination and primer–dimer artifact. BM = Bone marrow; AT = Adipose tissue; MSCs = Mesenchymal stem cells.

    Techniques Used: SYBR Green Assay, Reverse Transcription Polymerase Chain Reaction, Derivative Assay, Software, Fluorescence

    12) Product Images from "Antioxidant mechanism of black garlic extract involving nuclear factor erythroid 2-like factor 2 pathway"

    Article Title: Antioxidant mechanism of black garlic extract involving nuclear factor erythroid 2-like factor 2 pathway

    Journal: Nutrition Research and Practice

    doi: 10.4162/nrp.2017.11.3.206

    Effect of black garlic extract on mRNA expression of transcription factor and antioxidants in liver of rats. Rats were fed with 5 different diets for 5 weeks; NF, Normal fat (7% fat); HF, High-fat (20% fat + 1% cholesterol); HF+BGE 0.5, High-fat + 0.5% black garlic extract (BGE); HF+BGE 1.0, High-fat + 1.0% BGE; HF+BGE 1.5, High-fat + 1.5% BGE. Total RNA was isolated using TRI-reagent, and cDNA was synthesized using 3 µg of total RNA with SuperScript II reverse transcriptase. Real-time PCR was performed using SYBR green and standard procedures to assess the mRNA expression of the primer in liver samples obtained from each group. An Applied Biosystems StepOne softwere v2.1 was used. Each bar represents the mean ± SD of three independent experiments. Different letters above each bar indicate significant differences among the groups at α = 0.05 as determined by Duncan's multiple range tests. GR, glutathione reductase; NQO1, NAD(P)H:quinone-oxidoreductase-1; HO-1, heme oxygenase-1; Nrf2, nuclear factor erythroid 2-like factor; GSTA2, glutathione S-transferase alpha 2.
    Figure Legend Snippet: Effect of black garlic extract on mRNA expression of transcription factor and antioxidants in liver of rats. Rats were fed with 5 different diets for 5 weeks; NF, Normal fat (7% fat); HF, High-fat (20% fat + 1% cholesterol); HF+BGE 0.5, High-fat + 0.5% black garlic extract (BGE); HF+BGE 1.0, High-fat + 1.0% BGE; HF+BGE 1.5, High-fat + 1.5% BGE. Total RNA was isolated using TRI-reagent, and cDNA was synthesized using 3 µg of total RNA with SuperScript II reverse transcriptase. Real-time PCR was performed using SYBR green and standard procedures to assess the mRNA expression of the primer in liver samples obtained from each group. An Applied Biosystems StepOne softwere v2.1 was used. Each bar represents the mean ± SD of three independent experiments. Different letters above each bar indicate significant differences among the groups at α = 0.05 as determined by Duncan's multiple range tests. GR, glutathione reductase; NQO1, NAD(P)H:quinone-oxidoreductase-1; HO-1, heme oxygenase-1; Nrf2, nuclear factor erythroid 2-like factor; GSTA2, glutathione S-transferase alpha 2.

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

    13) Product Images from "Identification and Differential Expression of Two Thioredoxin h Isoforms in Germinating Seeds from Pea 1"

    Article Title: Identification and Differential Expression of Two Thioredoxin h Isoforms in Germinating Seeds from Pea 1

    Journal: Plant Physiology

    doi: 10.1104/pp.102.019562

    Expression of Trx h3 and h4 isoforms in seeds and seedlings by quantitative PCR. cDNAs produced by RT of total RNAs from cotyledons and embryo axes from seeds imbibed for 0 to 46 h (UG, ungerminated seeds; G, germinated seeds) and from green leaves (GL) and roots (R) from 7-d-old seedlings were used as templates in quantitative PCR realized in the presence of Sybr Green. Copy number of each isoform was determined using a standard curve established with the recombinant plasmid containing the corresponding full-length sequence.
    Figure Legend Snippet: Expression of Trx h3 and h4 isoforms in seeds and seedlings by quantitative PCR. cDNAs produced by RT of total RNAs from cotyledons and embryo axes from seeds imbibed for 0 to 46 h (UG, ungerminated seeds; G, germinated seeds) and from green leaves (GL) and roots (R) from 7-d-old seedlings were used as templates in quantitative PCR realized in the presence of Sybr Green. Copy number of each isoform was determined using a standard curve established with the recombinant plasmid containing the corresponding full-length sequence.

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction, Produced, SYBR Green Assay, Recombinant, Plasmid Preparation, Sequencing

    14) Product Images from "Epigenetic remodelling of gene expression profiles of neoplastic and normal tissues: immunotherapeutic implications"

    Article Title: Epigenetic remodelling of gene expression profiles of neoplastic and normal tissues: immunotherapeutic implications

    Journal: British Journal of Cancer

    doi: 10.1038/bjc.2012.361

    Quantitative RT–PCR analysis of P1A expression in tumour and normal tissues from 5-AZA-CdR-treated mice. Total RNA was extracted from tumour, bone marrow, blood, and spleen samples from 5-AZA-CdR-treated mice. Retrotranscribed RNA was subjected to SYBR Green quantitative real-time RT–PCR analysis using P1A- and β -actin-specific primers. Data are reported as P1A molecules/ β -actin molecules. Columns, mean values of P1A molecules/ β -actin molecules from six distinct mice; bars, s.d.; * P ⩽0.05 vs tumour samples.
    Figure Legend Snippet: Quantitative RT–PCR analysis of P1A expression in tumour and normal tissues from 5-AZA-CdR-treated mice. Total RNA was extracted from tumour, bone marrow, blood, and spleen samples from 5-AZA-CdR-treated mice. Retrotranscribed RNA was subjected to SYBR Green quantitative real-time RT–PCR analysis using P1A- and β -actin-specific primers. Data are reported as P1A molecules/ β -actin molecules. Columns, mean values of P1A molecules/ β -actin molecules from six distinct mice; bars, s.d.; * P ⩽0.05 vs tumour samples.

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

    15) Product Images from "The effect of fucoxanthin rich power on the lipid metabolism in rats with a high fat diet"

    Article Title: The effect of fucoxanthin rich power on the lipid metabolism in rats with a high fat diet

    Journal: Nutrition Research and Practice

    doi: 10.4162/nrp.2013.7.4.287

    Effect of fucoxanthin on mRNA expression of CPT1 in the liver of rats. Total RNA was isolated using TRI-reagenet and cDNA was synthesized using 3 µg of total RNA with SuperScript II reverse transcriptase. Realtime PCR was performed using SYBR green and standard procedures to assess the mRNA expression of primer in liver samples obtained from each group. An Applied Biosystem StepOne softwere v2.1 was used. Each bar represents the mean ± SD of three independent experiments. Different letters above each bar indicate significant differences among groups at α = 0.05 as determined by Duncan's multiple range test. CPT1, Carnitine palmitoyltransferase-1.
    Figure Legend Snippet: Effect of fucoxanthin on mRNA expression of CPT1 in the liver of rats. Total RNA was isolated using TRI-reagenet and cDNA was synthesized using 3 µg of total RNA with SuperScript II reverse transcriptase. Realtime PCR was performed using SYBR green and standard procedures to assess the mRNA expression of primer in liver samples obtained from each group. An Applied Biosystem StepOne softwere v2.1 was used. Each bar represents the mean ± SD of three independent experiments. Different letters above each bar indicate significant differences among groups at α = 0.05 as determined by Duncan's multiple range test. CPT1, Carnitine palmitoyltransferase-1.

    Techniques Used: Expressing, Isolation, Synthesized, Polymerase Chain Reaction, SYBR Green Assay

    Effect of fucoxanthin on mRNA expression of transcription factor and enzymes related to lipid metabolism in the liver of rats. Total RNA was isolated using TRI-reagenet and cDNA was synthesized using 3 µg of total RNA with SuperScript II reverse transcriptase. Realtime PCR was performed using SYBR green and standard procedures to assess the mRNA expression of primer in liver samples obtained from each group. An Applied Biosystem StepOne softwere v2.1 was used. Each bar represents the mean ± SD of three independent experiments. Different letters above each bar indicate significant differences among groups at α = 0.05 as determined by Duncan's multiple range test. SREBP-1C, Sterol regulatory element binding protein; G6PDH, Glucose-6-phosphate dehydrogenase; FAS, Fatty acid synthase; ACC, Acetyl-CoA carboxylase.
    Figure Legend Snippet: Effect of fucoxanthin on mRNA expression of transcription factor and enzymes related to lipid metabolism in the liver of rats. Total RNA was isolated using TRI-reagenet and cDNA was synthesized using 3 µg of total RNA with SuperScript II reverse transcriptase. Realtime PCR was performed using SYBR green and standard procedures to assess the mRNA expression of primer in liver samples obtained from each group. An Applied Biosystem StepOne softwere v2.1 was used. Each bar represents the mean ± SD of three independent experiments. Different letters above each bar indicate significant differences among groups at α = 0.05 as determined by Duncan's multiple range test. SREBP-1C, Sterol regulatory element binding protein; G6PDH, Glucose-6-phosphate dehydrogenase; FAS, Fatty acid synthase; ACC, Acetyl-CoA carboxylase.

    Techniques Used: Expressing, Isolation, Synthesized, Polymerase Chain Reaction, SYBR Green Assay, Binding Assay

    Effect of fucoxanthin on mRNA expression of enzymes related to cholesterol synthesis in the liver of rats. Total RNA was isolated using TRI-reagenet and cDNA was synthesized using 3 µg of total RNA with SuperScript II reverse transcriptase. Realtime PCR was performed using SYBR green and standard procedures to assess the mRNA expression of primer in liver samples obtained from each group. An Applied Biosystem StepOne softwere v2.1 was used. Each bar represents the mean ± SD of three independent experiments. Different letters above each bar indicate significant differences among groups at α = 0.05 as determined by Duncan's multiple range test. HMG-CoA, Hydroxy-3-methylglutaryl coenzyme A; ACAT, Acyl-CoA cholesterol acyltransferase; LCAT, lecithin-cholesterol acyltransferase.
    Figure Legend Snippet: Effect of fucoxanthin on mRNA expression of enzymes related to cholesterol synthesis in the liver of rats. Total RNA was isolated using TRI-reagenet and cDNA was synthesized using 3 µg of total RNA with SuperScript II reverse transcriptase. Realtime PCR was performed using SYBR green and standard procedures to assess the mRNA expression of primer in liver samples obtained from each group. An Applied Biosystem StepOne softwere v2.1 was used. Each bar represents the mean ± SD of three independent experiments. Different letters above each bar indicate significant differences among groups at α = 0.05 as determined by Duncan's multiple range test. HMG-CoA, Hydroxy-3-methylglutaryl coenzyme A; ACAT, Acyl-CoA cholesterol acyltransferase; LCAT, lecithin-cholesterol acyltransferase.

    Techniques Used: Expressing, Isolation, Synthesized, Polymerase Chain Reaction, SYBR Green Assay

    Effect of fucoxanthin on the mRNA expression of CYP7A1 in the liver of rats. Total RNA was isolated using TRI-reagenet and cDNA was synthesized using 3 µg of total RNA with SuperScript II reverse transcriptase. Realtime PCR was performed using SYBR green and standard procedures to assess the mRNA expression of primer in liver samples obtained from each group. An Applied Biosystem StepOne softwere v2.1 was used. Each bar represents the mean ± SD of three independent experiments. Different letters above each bar indicate significant differences among groups at α = 0.05 as determined by Duncan's multiple range test. CYP7A1, Cholesterol 7α-hydroxylase1.
    Figure Legend Snippet: Effect of fucoxanthin on the mRNA expression of CYP7A1 in the liver of rats. Total RNA was isolated using TRI-reagenet and cDNA was synthesized using 3 µg of total RNA with SuperScript II reverse transcriptase. Realtime PCR was performed using SYBR green and standard procedures to assess the mRNA expression of primer in liver samples obtained from each group. An Applied Biosystem StepOne softwere v2.1 was used. Each bar represents the mean ± SD of three independent experiments. Different letters above each bar indicate significant differences among groups at α = 0.05 as determined by Duncan's multiple range test. CYP7A1, Cholesterol 7α-hydroxylase1.

    Techniques Used: Expressing, Isolation, Synthesized, Polymerase Chain Reaction, SYBR Green Assay

    16) Product Images from "Distinctive profiles of tumor-infiltrating immune cells and association with intensity of infiltration in colorectal cancer"

    Article Title: Distinctive profiles of tumor-infiltrating immune cells and association with intensity of infiltration in colorectal cancer

    Journal: Oncology Letters

    doi: 10.3892/ol.2018.7771

    Analysis of the profile of infiltrating immune cells isolated from tumor and non-tumorous adjacent tissue. Immune cells were isolated from tissue blocks collected from selected patients with CRC during surgery via collagen IV digestion and gradient density centrifugation. Total RNA was extracted from the cells and subsequently reverse transcribed to cDNA. Specific primer sets were designed for transcription factors (A) Foxp3, (B) RORC, (C) BCL-6, (D) GATA-3 and (E) Tbx21 representing Treg, Th17, Tfh, Th2 and Th1 cells, respectively. qPCR was performed using the SYBR-Green method with specific primers to quantify the abundance of each subsets of infiltrating immune cells. GAPDH was amplified simultaneously for normalization. Data were analyzed using the 2 −ΔΔCq method and presented as relative values to GAPDH. T cells were isolated from tumor and non-tumorous adjacent tissue of 6 selected patients with CRC using T cell-specific microbeads. qPCR was performed on RNA isolated from T cells for quantification of (F) CXCL9, (G) CXCR3 and (H) CXCL10. Data are presented as the mean ± standard error of mean. Statistical analysis was performed using Student's t-test. *P
    Figure Legend Snippet: Analysis of the profile of infiltrating immune cells isolated from tumor and non-tumorous adjacent tissue. Immune cells were isolated from tissue blocks collected from selected patients with CRC during surgery via collagen IV digestion and gradient density centrifugation. Total RNA was extracted from the cells and subsequently reverse transcribed to cDNA. Specific primer sets were designed for transcription factors (A) Foxp3, (B) RORC, (C) BCL-6, (D) GATA-3 and (E) Tbx21 representing Treg, Th17, Tfh, Th2 and Th1 cells, respectively. qPCR was performed using the SYBR-Green method with specific primers to quantify the abundance of each subsets of infiltrating immune cells. GAPDH was amplified simultaneously for normalization. Data were analyzed using the 2 −ΔΔCq method and presented as relative values to GAPDH. T cells were isolated from tumor and non-tumorous adjacent tissue of 6 selected patients with CRC using T cell-specific microbeads. qPCR was performed on RNA isolated from T cells for quantification of (F) CXCL9, (G) CXCR3 and (H) CXCL10. Data are presented as the mean ± standard error of mean. Statistical analysis was performed using Student's t-test. *P

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

    17) Product Images from "Comparative Analysis of Cardiovascular Development Related Genes in Stem Cells Isolated from Deciduous Pulp and Adipose Tissue"

    Article Title: Comparative Analysis of Cardiovascular Development Related Genes in Stem Cells Isolated from Deciduous Pulp and Adipose Tissue

    Journal: The Scientific World Journal

    doi: 10.1155/2014/186508

    Transcription factors that are involved in cardiovascular-related development pathways in ASC and SHED. (a) Major pathways in ASC are development of blood vessel, vasculogenesis, angiogenesis, growth of blood vessel, cardiogenesis, endothelial cell development, and proliferation of cardiomyocytes. (b) Major pathways in SHED are development of blood vessel, vasculogenesis, cardiogenesis, angiogenesis, proliferation of cardiomyocytes and endothelial cells, and differentiation of cardiomyocytes. (c) Validation of gene expression by reverse transcriptase and real-time PCR. Reverse transcriptase of selected genes. mRNA expression of GATA6, RB1, NOTCH2, and MSX2 by real-time PCR using SYBR green reagent with values normalized to 18sRNA.
    Figure Legend Snippet: Transcription factors that are involved in cardiovascular-related development pathways in ASC and SHED. (a) Major pathways in ASC are development of blood vessel, vasculogenesis, angiogenesis, growth of blood vessel, cardiogenesis, endothelial cell development, and proliferation of cardiomyocytes. (b) Major pathways in SHED are development of blood vessel, vasculogenesis, cardiogenesis, angiogenesis, proliferation of cardiomyocytes and endothelial cells, and differentiation of cardiomyocytes. (c) Validation of gene expression by reverse transcriptase and real-time PCR. Reverse transcriptase of selected genes. mRNA expression of GATA6, RB1, NOTCH2, and MSX2 by real-time PCR using SYBR green reagent with values normalized to 18sRNA.

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

    18) Product Images from "Mechanism of Hepatitis C Virus (HCV)-induced Osteopontin and Its Role in Epithelial to Mesenchymal Transition of Hepatocytes *"

    Article Title: Mechanism of Hepatitis C Virus (HCV)-induced Osteopontin and Its Role in Epithelial to Mesenchymal Transition of Hepatocytes *

    Journal: The Journal of Biological Chemistry

    doi: 10.1074/jbc.M113.492314

    SYBR Green RT-PCR
    Figure Legend Snippet: SYBR Green RT-PCR

    Techniques Used: SYBR Green Assay, Reverse Transcription Polymerase Chain Reaction

    19) Product Images from "Epigenetic Changes in Mitochondrial Superoxide Dismutase in the Retina and the Development of Diabetic Retinopathy"

    Article Title: Epigenetic Changes in Mitochondrial Superoxide Dismutase in the Retina and the Development of Diabetic Retinopathy

    Journal: Diabetes

    doi: 10.2337/db10-0133

    H4K20me3 and acetyl H3K9 at sod2 , and evaluation of ChIP controls in retinal endothelial cells. A and B : H4K20me3 and acetyl H3K9 at the sod2 promoter and enhancer were measured by ChIP assay with SYBR green-based real-time q-PCR. Rabbit IgG served as a negative antibody control (indicated as ^). C : ChIP controls were verified by PCR. Crosslinked cells were immunoprecipitated with acetyl H3K9 or TFIIB antibody or normal rabbit IgG. The sod2 promoter occupied by acetyl H3K9 and the β-actin promoter occupied by TFIIB were amplified in purified ChIP-DNA. For the ChIP assay, the positive and negative controls were β-actin promoter occupied by TFIIB and the sod2 promoter occupied by IgG, respectively. The internal control included the input. 5 and 20, cells incubated in 5 mmol/L glucose or 20 mmol/L glucose; +Mn, cells transfected with sod2 plasmid, followed incubation in 20 mmol/L glucose for 4 days; and Mann, cells incubated in 20 mmol/L mannitol instead of 20 mmol/L glucose. * P
    Figure Legend Snippet: H4K20me3 and acetyl H3K9 at sod2 , and evaluation of ChIP controls in retinal endothelial cells. A and B : H4K20me3 and acetyl H3K9 at the sod2 promoter and enhancer were measured by ChIP assay with SYBR green-based real-time q-PCR. Rabbit IgG served as a negative antibody control (indicated as ^). C : ChIP controls were verified by PCR. Crosslinked cells were immunoprecipitated with acetyl H3K9 or TFIIB antibody or normal rabbit IgG. The sod2 promoter occupied by acetyl H3K9 and the β-actin promoter occupied by TFIIB were amplified in purified ChIP-DNA. For the ChIP assay, the positive and negative controls were β-actin promoter occupied by TFIIB and the sod2 promoter occupied by IgG, respectively. The internal control included the input. 5 and 20, cells incubated in 5 mmol/L glucose or 20 mmol/L glucose; +Mn, cells transfected with sod2 plasmid, followed incubation in 20 mmol/L glucose for 4 days; and Mann, cells incubated in 20 mmol/L mannitol instead of 20 mmol/L glucose. * P

    Techniques Used: Chromatin Immunoprecipitation, SYBR Green Assay, Polymerase Chain Reaction, Immunoprecipitation, Amplification, Purification, Incubation, Transfection, Plasmid Preparation

    20) Product Images from "MicroRNAs activate natural killer cells through Toll-like receptor signaling"

    Article Title: MicroRNAs activate natural killer cells through Toll-like receptor signaling

    Journal: Blood

    doi: 10.1182/blood-2012-07-441360

    Downregulation of the NF-κB signaling pathway components in NK cells from lymphoma patients. (A,B) NK cells were isolated from PBMCs of both healthy donors and lymphoma patients as described in the Materials and Methods section. The purified NK cells were then immediately subjected to RNA extraction and cDNA synthesis. The expression levels of p65 (A) and IRAK1 (B) were determined by SYBR Green real-time RT-PCR assay.
    Figure Legend Snippet: Downregulation of the NF-κB signaling pathway components in NK cells from lymphoma patients. (A,B) NK cells were isolated from PBMCs of both healthy donors and lymphoma patients as described in the Materials and Methods section. The purified NK cells were then immediately subjected to RNA extraction and cDNA synthesis. The expression levels of p65 (A) and IRAK1 (B) were determined by SYBR Green real-time RT-PCR assay.

    Techniques Used: Isolation, Purification, RNA Extraction, Expressing, SYBR Green Assay, Quantitative RT-PCR

    21) Product Images from "Impact of gestational low-protein intake on embryonic kidney microRNA expression and in the nephron progenitor cells of the male offspring fetus"

    Article Title: Impact of gestational low-protein intake on embryonic kidney microRNA expression and in the nephron progenitor cells of the male offspring fetus

    Journal: bioRxiv

    doi: 10.1101/2020.02.18.954800

    Expression of mRNA estimated by SyBR green RT-qPCR of metanephros from the 17th day LP fetus. The expression was normalized with GAPDH. Data are expressed as fold change (mean ± SD, n = 4) concerning the control group. * p≤0.05: statistical significance versus NP.
    Figure Legend Snippet: Expression of mRNA estimated by SyBR green RT-qPCR of metanephros from the 17th day LP fetus. The expression was normalized with GAPDH. Data are expressed as fold change (mean ± SD, n = 4) concerning the control group. * p≤0.05: statistical significance versus NP.

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

    22) Product Images from "The Aryl Hydrocarbon Receptor Regulates an Essential Transcriptional Element in the Immunoglobulin Heavy Chain Gene"

    Article Title: The Aryl Hydrocarbon Receptor Regulates an Essential Transcriptional Element in the Immunoglobulin Heavy Chain Gene

    Journal: Cellular immunology

    doi: 10.1016/j.cellimm.2015.02.012

    AhR knockdown and a competitive AhR antagonist decrease TCDD-induced Cyp1A1 expression confirming disruption of the AhR signaling pathway. “Wild Type” (WT) refers to a variant of the CH12.LX murine B-cell line that endogenously expresses IgA and has a stably integrated 3′ Igh RR-regulated γ2b transgene (CH12.γ2b-3′ Igh RR cells). “shAhR” denotes CH12.γ2b-3′ Igh RR cells that stably express shRNA to the AhR via lentiviral-mediated delivery of one of two different shAhR constructs (denoted shAhR11 or shAhR12). A) Whole cell lysates were collected from WT and shAhR cells and 20 μg of protein was subjected to western blot analysis and probed for the AhR or the loading control β-actin. B) Cells were either treated with 10 nM TCDD or the vehicle control, 0.01% DMSO (not shown), for 8 hours. For the AhR antagonist treatment (first data set), WT cells were either pretreated with 30 μM of CH-223191 (denoted “AhRA”) or the vehicle control for CH-223191 (0.1% DMSO) for one hour prior to the 8 hr treatment with 10 nM TCDD; the final DMSO concentration was 0.11%. Total RNA was extracted, one μg was reverse transcribed to cDNA, and 5 ng of cDNA was used to amplify Cyp1A1 and β-actin via SYBR Green quantitative real-time PCR. The ΔΔ Ct method was used to determine the Cyp1A1 levels as a fold-difference or relative quantification (RQ) compared to the highest Cyp1A1-inducing treatment group (WT cells treated with 10 nM TCDD). NA and DMSO treatments caused no appreciable expression of Cyp1A1 (data not shown). Results are representative of at least two separate experiments (n=3 for each treatment group). Statistical significance was determined by a one-tailed t-test. “***” and “**” denote significance from the DMSO control at p
    Figure Legend Snippet: AhR knockdown and a competitive AhR antagonist decrease TCDD-induced Cyp1A1 expression confirming disruption of the AhR signaling pathway. “Wild Type” (WT) refers to a variant of the CH12.LX murine B-cell line that endogenously expresses IgA and has a stably integrated 3′ Igh RR-regulated γ2b transgene (CH12.γ2b-3′ Igh RR cells). “shAhR” denotes CH12.γ2b-3′ Igh RR cells that stably express shRNA to the AhR via lentiviral-mediated delivery of one of two different shAhR constructs (denoted shAhR11 or shAhR12). A) Whole cell lysates were collected from WT and shAhR cells and 20 μg of protein was subjected to western blot analysis and probed for the AhR or the loading control β-actin. B) Cells were either treated with 10 nM TCDD or the vehicle control, 0.01% DMSO (not shown), for 8 hours. For the AhR antagonist treatment (first data set), WT cells were either pretreated with 30 μM of CH-223191 (denoted “AhRA”) or the vehicle control for CH-223191 (0.1% DMSO) for one hour prior to the 8 hr treatment with 10 nM TCDD; the final DMSO concentration was 0.11%. Total RNA was extracted, one μg was reverse transcribed to cDNA, and 5 ng of cDNA was used to amplify Cyp1A1 and β-actin via SYBR Green quantitative real-time PCR. The ΔΔ Ct method was used to determine the Cyp1A1 levels as a fold-difference or relative quantification (RQ) compared to the highest Cyp1A1-inducing treatment group (WT cells treated with 10 nM TCDD). NA and DMSO treatments caused no appreciable expression of Cyp1A1 (data not shown). Results are representative of at least two separate experiments (n=3 for each treatment group). Statistical significance was determined by a one-tailed t-test. “***” and “**” denote significance from the DMSO control at p

    Techniques Used: Expressing, Variant Assay, Stable Transfection, shRNA, Construct, Western Blot, Concentration Assay, SYBR Green Assay, Real-time Polymerase Chain Reaction, One-tailed Test

    23) Product Images from "A Transcriptionally Permissive Epigenetic Landscape at the Vasoactive Intestinal Peptide Receptor-1 Promoter Suggests an Euchromatin Nuclear Position in Murine CD4 T Cells"

    Article Title: A Transcriptionally Permissive Epigenetic Landscape at the Vasoactive Intestinal Peptide Receptor-1 Promoter Suggests an Euchromatin Nuclear Position in Murine CD4 T Cells

    Journal: Regulatory peptides

    doi: 10.1016/j.regpep.2009.08.010

    Differential enrichment of H3K9/14ac and H3K4me3 histone modifications at the VPAC1 promoter A. Flow cytometric analysis of mouse CD4 + T cells and CD45R + B cells. Left Panel : Non-adherent splenocytes were stained with anti-CD4-FITC pre- (red line) and post- (blue line) CD4 positive magnetic bead separation ( Materials and Methods ) with typical purities ≥93% (n=3). Right Panel : CD4 depleted non-adherent splenocytes were subsequently stained pre- (red line) and post- (blue line) CD45R-FITC positive magnetic bead separation ( Materials and Methods ) with typical purities ≥97% (n=3). B. Bar graph for Taqman qPCR measurements for VPAC1 from indicated cell type. A hash mark indicates a shift in the y-axis. Data is represented as means +/- SEM for relative VPAC1 levels normalized to HPRT and calculated by 2 (-ΔCt) formula from three independent experiments. The asterisk (*) represents a p≤ 0.05 as compared to CD4 + T cells. C-D. ChIP analysis was performed using purified CD4 + T and CD45R + B cells followed by quantitative SYBR green PCR using VPAC1 primer set 2 or genomic negative control. C. Amplification reactions from SYBR green qPCR using primer set 2 were separated on a 3% agarose gel, stained with ethidium bromide and visualized by a CCD camera (Syngene). This experiment was repeated twice with similar results. D. Bar graph showing fold-enrichment over IgG control of H3K9/14ac and H3K4me3 from the indicated cell types. Cycle thresholds (Ct) were subtracted from input Ct values to obtain relative ΔCt values. Fold increases were calculated by 2 -(ΔCt) formula with non specific IgG levels normalized to input arbitrarily set to 1 (not shown). Data is presented as means +/- SEM from three biologically independent experiments (*, p≤0.05).
    Figure Legend Snippet: Differential enrichment of H3K9/14ac and H3K4me3 histone modifications at the VPAC1 promoter A. Flow cytometric analysis of mouse CD4 + T cells and CD45R + B cells. Left Panel : Non-adherent splenocytes were stained with anti-CD4-FITC pre- (red line) and post- (blue line) CD4 positive magnetic bead separation ( Materials and Methods ) with typical purities ≥93% (n=3). Right Panel : CD4 depleted non-adherent splenocytes were subsequently stained pre- (red line) and post- (blue line) CD45R-FITC positive magnetic bead separation ( Materials and Methods ) with typical purities ≥97% (n=3). B. Bar graph for Taqman qPCR measurements for VPAC1 from indicated cell type. A hash mark indicates a shift in the y-axis. Data is represented as means +/- SEM for relative VPAC1 levels normalized to HPRT and calculated by 2 (-ΔCt) formula from three independent experiments. The asterisk (*) represents a p≤ 0.05 as compared to CD4 + T cells. C-D. ChIP analysis was performed using purified CD4 + T and CD45R + B cells followed by quantitative SYBR green PCR using VPAC1 primer set 2 or genomic negative control. C. Amplification reactions from SYBR green qPCR using primer set 2 were separated on a 3% agarose gel, stained with ethidium bromide and visualized by a CCD camera (Syngene). This experiment was repeated twice with similar results. D. Bar graph showing fold-enrichment over IgG control of H3K9/14ac and H3K4me3 from the indicated cell types. Cycle thresholds (Ct) were subtracted from input Ct values to obtain relative ΔCt values. Fold increases were calculated by 2 -(ΔCt) formula with non specific IgG levels normalized to input arbitrarily set to 1 (not shown). Data is presented as means +/- SEM from three biologically independent experiments (*, p≤0.05).

    Techniques Used: Flow Cytometry, Staining, Real-time Polymerase Chain Reaction, Chromatin Immunoprecipitation, Purification, SYBR Green Assay, Polymerase Chain Reaction, Negative Control, Amplification, Agarose Gel Electrophoresis

    24) Product Images from "Wolbachia restricts insect-specific flavivirus infection in Aedes aegypti cells"

    Article Title: Wolbachia restricts insect-specific flavivirus infection in Aedes aegypti cells

    Journal: The Journal of General Virology

    doi: 10.1099/jgv.0.000617

    Effect of w MelPop on CFAV and PCLV infection in Aag2 cells. (a) Detection of CFAV, Wolbachia or PCLV in Aag2, Aag2 w MelPop and two different cultures of Aag2 w MelPop cells treated with tetracycline (Aag2 w MelPop-tet sets 1 and 2) cells by RT-PCR. Actin was used as loading control. (b) Detection of CFAV or PCLV in C6/36 cells incubated with supernatant of Aag2, Aag2 w MelPop or Aag2 w MelPop treated with tetracycline (two different cultures, Aag2 w MelPop-tet sets 1 and 2) by RT-PCR. Actin was used as a loading control. (c) Quantification of CFAV RNA in Aag2 w MelPop (Wol) or Aag2 w MelPop treated with tetracycline (Tet) cells after incubation with Aag2 supernatant containing CFAV by SYBR Green. S7 was used as internal control. Relative RNA expression is represented as (CFAV/S7). Error bars show sem from three independent experiments. (d) Quantification of PCLV RNA in Aag2 w MelPop (Wol) or Aag2 w MelPop treated with tetracycline (Tet) cells, either after incubation with Aag2 supernatant harbouring PCLV or untreated by SYBR Green. S7 was used as an internal control. Relative RNA expression is represented as (PCLV/S7) and mock-infected tetracycline cells were set to 1. Error bars show sem from three independent experiments. * P ≤0.05.
    Figure Legend Snippet: Effect of w MelPop on CFAV and PCLV infection in Aag2 cells. (a) Detection of CFAV, Wolbachia or PCLV in Aag2, Aag2 w MelPop and two different cultures of Aag2 w MelPop cells treated with tetracycline (Aag2 w MelPop-tet sets 1 and 2) cells by RT-PCR. Actin was used as loading control. (b) Detection of CFAV or PCLV in C6/36 cells incubated with supernatant of Aag2, Aag2 w MelPop or Aag2 w MelPop treated with tetracycline (two different cultures, Aag2 w MelPop-tet sets 1 and 2) by RT-PCR. Actin was used as a loading control. (c) Quantification of CFAV RNA in Aag2 w MelPop (Wol) or Aag2 w MelPop treated with tetracycline (Tet) cells after incubation with Aag2 supernatant containing CFAV by SYBR Green. S7 was used as internal control. Relative RNA expression is represented as (CFAV/S7). Error bars show sem from three independent experiments. (d) Quantification of PCLV RNA in Aag2 w MelPop (Wol) or Aag2 w MelPop treated with tetracycline (Tet) cells, either after incubation with Aag2 supernatant harbouring PCLV or untreated by SYBR Green. S7 was used as an internal control. Relative RNA expression is represented as (PCLV/S7) and mock-infected tetracycline cells were set to 1. Error bars show sem from three independent experiments. * P ≤0.05.

    Techniques Used: Infection, Reverse Transcription Polymerase Chain Reaction, Incubation, SYBR Green Assay, RNA Expression

    25) Product Images from "Blood Metallothionein Transcript as a Biomarker for Metal Sensitivity: Low Blood Metallothionein Transcripts in Arsenicosis Patients from Guizhou, China"

    Article Title: Blood Metallothionein Transcript as a Biomarker for Metal Sensitivity: Low Blood Metallothionein Transcripts in Arsenicosis Patients from Guizhou, China

    Journal: Environmental Health Perspectives

    doi: 10.1289/ehp.10035

    The sex difference in human MT-1A and MT-2A expression in blood cells of arsenicosis patients (26 males and 22 females) and healthy subjects (23 males and 25 females) in Guizhou, China. Total RNA was extracted, purified, reverse-transcripted, and subjected to SYBR Green real-time quantitative RT-PCR with MT isoform primers. Data are mean ± SE.
    Figure Legend Snippet: The sex difference in human MT-1A and MT-2A expression in blood cells of arsenicosis patients (26 males and 22 females) and healthy subjects (23 males and 25 females) in Guizhou, China. Total RNA was extracted, purified, reverse-transcripted, and subjected to SYBR Green real-time quantitative RT-PCR with MT isoform primers. Data are mean ± SE.

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

    The expression of human MT isoforms in buccal cells of arsenic-exposed patients ( n = 44) and healthy subjects ( n = 12) in Guizhou, China. Total RNA was extracted, purified, reverse-transcripted, and subjected to SYBR Green real-time quantitative RT-PCR with MT isoform primers. Data are mean ± SE. *Significantly different from controls at p
    Figure Legend Snippet: The expression of human MT isoforms in buccal cells of arsenic-exposed patients ( n = 44) and healthy subjects ( n = 12) in Guizhou, China. Total RNA was extracted, purified, reverse-transcripted, and subjected to SYBR Green real-time quantitative RT-PCR with MT isoform primers. Data are mean ± SE. *Significantly different from controls at p

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

    The expression of human MT isoforms in blood cells of arsenic-exposed patients ( n = 48) and healthy subjects ( n = 48) in Guizhou, China. Total RNA was extracted, purified, reverse-transcripted, and subjected to SYBR Green real-time quantitative RT-PCR with MT isoform primers. Data are mean ± SE. *Significantly different from healthy controls at p
    Figure Legend Snippet: The expression of human MT isoforms in blood cells of arsenic-exposed patients ( n = 48) and healthy subjects ( n = 48) in Guizhou, China. Total RNA was extracted, purified, reverse-transcripted, and subjected to SYBR Green real-time quantitative RT-PCR with MT isoform primers. Data are mean ± SE. *Significantly different from healthy controls at p

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

    Related Articles

    SYBR Green Assay:

    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
    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.

    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. ..

    Article Title: Characterization and use of a rabbit-anti-mouse VPAC1 antibody by flow cytometry
    Article Snippet: .. Fast SYBR green PCR master mix was purchased from Applied Biosystems (Foster City, CA). .. VIP was purchased from American Peptide (Sunnyvale, CA) and Cyclic AMP EIA Kit from Cayman Chemical Company (Ann Arbor, MI).

    Article Title: Coordination of Cell Proliferation and Cell Fate Determination by CES-1 Snail
    Article Snippet: .. Quantitative real-time PCR (qPCR) Fast SYBR green master mix (Applied Biosystems) was used to amplify cDNA templates by real-time PCR. .. Each sample was performed in triplicate on a Biorad CFX96 real-time PCR machine.

    Article Title: NFAT2 Regulates Generation of Innate-Like CD8+ T Lymphocytes and CD8+ T Lymphocytes Responses
    Article Snippet: .. Real-time polymerase chain reactions for NFAT2 and PLZF were performed using SYBR Green master mix (Applied Biosystems) and for Blimp-1, Tbx21, IL-4, Eomes using Taqman probes. ..

    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
    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.

    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. ..

    Article Title: Coordination of Cell Proliferation and Cell Fate Determination by CES-1 Snail
    Article Snippet: .. Quantitative real-time PCR (qPCR) Fast SYBR green master mix (Applied Biosystems) was used to amplify cDNA templates by real-time PCR. .. Each sample was performed in triplicate on a Biorad CFX96 real-time PCR machine.

    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.

    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.

    Article Title: Characterization and use of a rabbit-anti-mouse VPAC1 antibody by flow cytometry
    Article Snippet: .. Fast SYBR green PCR master mix was purchased from Applied Biosystems (Foster City, CA). .. VIP was purchased from American Peptide (Sunnyvale, CA) and Cyclic AMP EIA Kit from Cayman Chemical Company (Ann Arbor, MI).

    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.

    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.

    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. ..

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 90
    Thermo Fisher sybr green i
    Correlation of detection of virus by Bio-Plex and RT-qPCR. Correlation between qPCR (Ct value) and Bio-Plex (MFI) detection of in-vitro grown ISAV standard curve by (A) <t>SYBR</t> <t>GREEN</t> I and (B)TaqMan; Correlation of detection of copy number of ISAV in plasma samples by (C) SYBR Green I (n = 13) and (D) TaqMan chemistries (n = 8).
    Sybr Green I, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 13018 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sybr green i/product/Thermo Fisher
    Average 90 stars, based on 13018 article reviews
    Price from $9.99 to $1999.99
    sybr green i - by Bioz Stars, 2020-07
    90/100 stars
      Buy from Supplier

    99
    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
    https://www.bioz.com/result/fast sybr green master mix/product/Thermo Fisher
    Average 99 stars, based on 3358 article reviews
    Price from $9.99 to $1999.99
    fast sybr green master mix - by Bioz Stars, 2020-07
    99/100 stars
      Buy from Supplier

    99
    Thermo Fisher real time quantitative pcr analysis
    Confirmation of RNA and protein expression levels by real-time quantitative <t>PCR</t> and western blotting analysis (n = 3, per treatment type). (A) The expression of LASP1 and ferritin light and heavy chain mRNAs relative to that of GAPDH, the internal control, as detected by real-time PCR. (B) Expression of LASP1 and ferritin light and heavy chains, as assessed by western blotting analysis. The signal corresponding to the protein bands was quantified using densitometric scanning and the relative protein abundance was determined after normalization with the levels of β-actin protein. Data are presented as mean ± SD and compared using the t -test for independent samples. * p
    Real Time Quantitative Pcr Analysis, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 266 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/real time quantitative pcr analysis/product/Thermo Fisher
    Average 99 stars, based on 266 article reviews
    Price from $9.99 to $1999.99
    real time quantitative pcr analysis - by Bioz Stars, 2020-07
    99/100 stars
      Buy from Supplier

    Image Search Results


    Correlation of detection of virus by Bio-Plex and RT-qPCR. Correlation between qPCR (Ct value) and Bio-Plex (MFI) detection of in-vitro grown ISAV standard curve by (A) SYBR GREEN I and (B)TaqMan; Correlation of detection of copy number of ISAV in plasma samples by (C) SYBR Green I (n = 13) and (D) TaqMan chemistries (n = 8).

    Journal: PLoS ONE

    Article Title: Development, Characterisation and Application of Monoclonal Antibodies for the Detection and Quantification of Infectious Salmon Anaemia Virus in Plasma Samples Using Luminex Bead Array Technology

    doi: 10.1371/journal.pone.0159155

    Figure Lengend Snippet: Correlation of detection of virus by Bio-Plex and RT-qPCR. Correlation between qPCR (Ct value) and Bio-Plex (MFI) detection of in-vitro grown ISAV standard curve by (A) SYBR GREEN I and (B)TaqMan; Correlation of detection of copy number of ISAV in plasma samples by (C) SYBR Green I (n = 13) and (D) TaqMan chemistries (n = 8).

    Article Snippet: 2.3.1 qRT-PCR with SYBR Green I chemistry or TaqMan® chemistry Real time PCR was performed on first strand cDNA (reverse transcribed from in-vitro transcribed RNA) using the Eppendorf® RealPlex2 Mastercycler gradient S instrument, using ISAV Segment 8 primer pair ISAV8F/ISAV8R (amplifying a 104 bp product), [ ], either with SYBR® Green I (Thermo Scientific) master mix or with TaqMan® master mix.

    Techniques: Quantitative RT-PCR, Real-time Polymerase Chain Reaction, In Vitro, SYBR Green Assay

    RT-qPCR standard curves. Standard curve relating viral copy number to Ct value from RT-Polymerase chain reaction using 10-fold dilutions of in-vitro transcribed RNA using (a) SYBR ® Green I and (b) TaqMan chemistries.

    Journal: PLoS ONE

    Article Title: Development, Characterisation and Application of Monoclonal Antibodies for the Detection and Quantification of Infectious Salmon Anaemia Virus in Plasma Samples Using Luminex Bead Array Technology

    doi: 10.1371/journal.pone.0159155

    Figure Lengend Snippet: RT-qPCR standard curves. Standard curve relating viral copy number to Ct value from RT-Polymerase chain reaction using 10-fold dilutions of in-vitro transcribed RNA using (a) SYBR ® Green I and (b) TaqMan chemistries.

    Article Snippet: 2.3.1 qRT-PCR with SYBR Green I chemistry or TaqMan® chemistry Real time PCR was performed on first strand cDNA (reverse transcribed from in-vitro transcribed RNA) using the Eppendorf® RealPlex2 Mastercycler gradient S instrument, using ISAV Segment 8 primer pair ISAV8F/ISAV8R (amplifying a 104 bp product), [ ], either with SYBR® Green I (Thermo Scientific) master mix or with TaqMan® master mix.

    Techniques: Quantitative RT-PCR, Polymerase Chain Reaction, In Vitro, SYBR Green Assay

    Silencing of Snail in Caki-2 by a lentivirus-delivered short inhibitory RNA. A. Map of the lentivirualRNAi vectors [pCMV-G NR-U6-shRNA]. U6, polymerase III promoter to drive the transcription of shRNAs; GFP (green fluorescent protein) were driven by CMV promoter. shRNAs were inserted into HindIII and BamHI sites. B. Strategy plan for generating Snail shRNA constructs. pCMV-G NR-U6 vector was linearized by HindIII and BamH1, and then designed shRNAs were inserted into this site. The positive colonies were screened by colony PCR using the primers flanking the inserted site of shRNA. Selected Colonies were finally confirmed by sequencing. C. Quantitative real-time RT-PCR compare the knock-down efficiency of 3 Snail shRNAs in Caki-2 cells. RNAs were isolated from cells and gene reverse transcription was performed. Quantitative PCR was performed by using SYBR Green PCR Master Mix. Data were normalized to GAPDH mRNA levels, by using efficiency (2 -ΔΔCt ) method. The data were analyzed by student t -test, and were presented as the means ± s.e.m (n = 3). D. Western blotting assay confirmed that effectiveness of Snail shRNA viruses on protein level.The intensity of bands were determined by Image J, and ratio of Snail to GAPDH were shown here.

    Journal: International Journal of Clinical and Experimental Pathology

    Article Title: Synergistic effects of snail and quercetin on renal cell carcinoma Caki-2 by altering AKT/mTOR/ERK1/2 signaling pathways

    doi:

    Figure Lengend Snippet: Silencing of Snail in Caki-2 by a lentivirus-delivered short inhibitory RNA. A. Map of the lentivirualRNAi vectors [pCMV-G NR-U6-shRNA]. U6, polymerase III promoter to drive the transcription of shRNAs; GFP (green fluorescent protein) were driven by CMV promoter. shRNAs were inserted into HindIII and BamHI sites. B. Strategy plan for generating Snail shRNA constructs. pCMV-G NR-U6 vector was linearized by HindIII and BamH1, and then designed shRNAs were inserted into this site. The positive colonies were screened by colony PCR using the primers flanking the inserted site of shRNA. Selected Colonies were finally confirmed by sequencing. C. Quantitative real-time RT-PCR compare the knock-down efficiency of 3 Snail shRNAs in Caki-2 cells. RNAs were isolated from cells and gene reverse transcription was performed. Quantitative PCR was performed by using SYBR Green PCR Master Mix. Data were normalized to GAPDH mRNA levels, by using efficiency (2 -ΔΔCt ) method. The data were analyzed by student t -test, and were presented as the means ± s.e.m (n = 3). D. Western blotting assay confirmed that effectiveness of Snail shRNA viruses on protein level.The intensity of bands were determined by Image J, and ratio of Snail to GAPDH were shown here.

    Article Snippet: Quantitative PCR was performed with SYBR Green PCR Master Mix (Thermo, F-415XL) on Applied Biosystems 7300 Fast Real-Time PCR System.

    Techniques: shRNA, Construct, Plasmid Preparation, Polymerase Chain Reaction, Sequencing, Quantitative RT-PCR, Isolation, Real-time Polymerase Chain Reaction, SYBR Green Assay, Western Blot

    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

    Confirmation of RNA and protein expression levels by real-time quantitative PCR and western blotting analysis (n = 3, per treatment type). (A) The expression of LASP1 and ferritin light and heavy chain mRNAs relative to that of GAPDH, the internal control, as detected by real-time PCR. (B) Expression of LASP1 and ferritin light and heavy chains, as assessed by western blotting analysis. The signal corresponding to the protein bands was quantified using densitometric scanning and the relative protein abundance was determined after normalization with the levels of β-actin protein. Data are presented as mean ± SD and compared using the t -test for independent samples. * p

    Journal: Proteome Science

    Article Title: Proteomic analysis on effectors involved in BMP-2-induced osteogenic differentiation of beagle bone marrow mesenchymal stem cells

    doi: 10.1186/1477-5956-12-13

    Figure Lengend Snippet: Confirmation of RNA and protein expression levels by real-time quantitative PCR and western blotting analysis (n = 3, per treatment type). (A) The expression of LASP1 and ferritin light and heavy chain mRNAs relative to that of GAPDH, the internal control, as detected by real-time PCR. (B) Expression of LASP1 and ferritin light and heavy chains, as assessed by western blotting analysis. The signal corresponding to the protein bands was quantified using densitometric scanning and the relative protein abundance was determined after normalization with the levels of β-actin protein. Data are presented as mean ± SD and compared using the t -test for independent samples. * p

    Article Snippet: The cDNA was then used as the template for real-time quantitative PCR analysis (Maxima SYBR Green/ROX qPCR Master Mix; Thermo Scientific, Pittsburgh, PA).

    Techniques: Expressing, Real-time Polymerase Chain Reaction, Western Blot