sybr green master mix Thermo Fisher Search Results


  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    Thermo Fisher sybr master mix
    HEV induces <t>ISG15</t> both in vitro and in vivo . (A) Replication kinetics of viral RNAs from the HEV P6 infectious cDNA clone and HEV P6GLuc replicon in Huh7-S10-3 liver cells. The cells were transfected with capped RNA transcripts from the genotype 3 HEV P6 infectious cDNA clone (HEV P6) or with the HEV P6GLuc replicon (HEV P6GLuc; P6 encoding Gaussia luciferase clone). The culture supernatants were collected at various time points and used to measure viral RNA replication levels by HEV real-time quantitative RT-PCR or by the Gaussia luciferase assay (GLuc assay). RLU, relative light units. (B) ISG15 mRNA expression levels in Huh7-S10-3 liver cells transfected with capped RNA transcripts from the HEV P6 infectious cDNA clone or the HEV P6GLuc replicon at 1 and 5 days posttransfection (dpt) were determined by <t>Sybr</t> green-based quantitative PCR (qPCR). The ISG15-mRNA fold change was calculated using the 2 −ΔΔ CT method for comparisons to untransfected control cells, and the RPS18 gene was used as the housekeeping gene. (C and D) ISG15 protein expression levels in Huh7-S10-3 cells transfected with capped RNA from the HEV P6 infectious cDNA clone or the HEV P6GLuc replicon (C and D) or the HEV P6 replication-deficient infectious cDNA clone (P6GAD) (D). CC, untransfected cell control. The ISG15 protein expression, at 5 days posttransfection, in cell lysates (40 μg/lane) was analyzed by Western blotting using anti-ISG15 antibody (1:1,000 dilution). Fold change in band density, as calculated via normalization of ISG15 with β-actin (loading control), was measured using ImageJ software (NIH, Bethesda, MD). (E) ISG15 and USP18 mRNA expression levels in swine liver tissue samples obtained from animals experimentally infected with HEV ( n = 3) at 3 weeks postinfection. The fold changes in swine ISG15 and USP18 mRNA levels were calculated using the 2 −ΔΔ CT method compared to the uninfected control animals ( n = 2), and the RPS18 gene was used as the housekeeping gene. The data represent means ± standard errors of the means (SEM) of results from three independent transfection experiments (A, B, and C), one experiment representative of two independent transfection experiments (D), and HEV-infected animals ( n = 3) compared to uninfected animals ( n = 2) (E). **, P ≤ 0.01.
    Sybr Master Mix, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 2195 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sybr master mix/product/Thermo Fisher
    Average 99 stars, based on 2195 article reviews
    Price from $9.99 to $1999.99
    sybr master mix - by Bioz Stars, 2020-07
    99/100 stars
      Buy from Supplier

    84
    Thermo Fisher powerup sybr green master mix thermo fisher scientific
    HEV induces <t>ISG15</t> both in vitro and in vivo . (A) Replication kinetics of viral RNAs from the HEV P6 infectious cDNA clone and HEV P6GLuc replicon in Huh7-S10-3 liver cells. The cells were transfected with capped RNA transcripts from the genotype 3 HEV P6 infectious cDNA clone (HEV P6) or with the HEV P6GLuc replicon (HEV P6GLuc; P6 encoding Gaussia luciferase clone). The culture supernatants were collected at various time points and used to measure viral RNA replication levels by HEV real-time quantitative RT-PCR or by the Gaussia luciferase assay (GLuc assay). RLU, relative light units. (B) ISG15 mRNA expression levels in Huh7-S10-3 liver cells transfected with capped RNA transcripts from the HEV P6 infectious cDNA clone or the HEV P6GLuc replicon at 1 and 5 days posttransfection (dpt) were determined by <t>Sybr</t> green-based quantitative PCR (qPCR). The ISG15-mRNA fold change was calculated using the 2 −ΔΔ CT method for comparisons to untransfected control cells, and the RPS18 gene was used as the housekeeping gene. (C and D) ISG15 protein expression levels in Huh7-S10-3 cells transfected with capped RNA from the HEV P6 infectious cDNA clone or the HEV P6GLuc replicon (C and D) or the HEV P6 replication-deficient infectious cDNA clone (P6GAD) (D). CC, untransfected cell control. The ISG15 protein expression, at 5 days posttransfection, in cell lysates (40 μg/lane) was analyzed by Western blotting using anti-ISG15 antibody (1:1,000 dilution). Fold change in band density, as calculated via normalization of ISG15 with β-actin (loading control), was measured using ImageJ software (NIH, Bethesda, MD). (E) ISG15 and USP18 mRNA expression levels in swine liver tissue samples obtained from animals experimentally infected with HEV ( n = 3) at 3 weeks postinfection. The fold changes in swine ISG15 and USP18 mRNA levels were calculated using the 2 −ΔΔ CT method compared to the uninfected control animals ( n = 2), and the RPS18 gene was used as the housekeeping gene. The data represent means ± standard errors of the means (SEM) of results from three independent transfection experiments (A, B, and C), one experiment representative of two independent transfection experiments (D), and HEV-infected animals ( n = 3) compared to uninfected animals ( n = 2) (E). **, P ≤ 0.01.
    Powerup Sybr Green Master Mix Thermo Fisher Scientific, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 84/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/powerup sybr green master mix thermo fisher scientific/product/Thermo Fisher
    Average 84 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    powerup sybr green master mix thermo fisher scientific - by Bioz Stars, 2020-07
    84/100 stars
      Buy from Supplier

    84
    Thermo Fisher sybr green master mix thermo fisher scientific cat
    HEV induces <t>ISG15</t> both in vitro and in vivo . (A) Replication kinetics of viral RNAs from the HEV P6 infectious cDNA clone and HEV P6GLuc replicon in Huh7-S10-3 liver cells. The cells were transfected with capped RNA transcripts from the genotype 3 HEV P6 infectious cDNA clone (HEV P6) or with the HEV P6GLuc replicon (HEV P6GLuc; P6 encoding Gaussia luciferase clone). The culture supernatants were collected at various time points and used to measure viral RNA replication levels by HEV real-time quantitative RT-PCR or by the Gaussia luciferase assay (GLuc assay). RLU, relative light units. (B) ISG15 mRNA expression levels in Huh7-S10-3 liver cells transfected with capped RNA transcripts from the HEV P6 infectious cDNA clone or the HEV P6GLuc replicon at 1 and 5 days posttransfection (dpt) were determined by <t>Sybr</t> green-based quantitative PCR (qPCR). The ISG15-mRNA fold change was calculated using the 2 −ΔΔ CT method for comparisons to untransfected control cells, and the RPS18 gene was used as the housekeeping gene. (C and D) ISG15 protein expression levels in Huh7-S10-3 cells transfected with capped RNA from the HEV P6 infectious cDNA clone or the HEV P6GLuc replicon (C and D) or the HEV P6 replication-deficient infectious cDNA clone (P6GAD) (D). CC, untransfected cell control. The ISG15 protein expression, at 5 days posttransfection, in cell lysates (40 μg/lane) was analyzed by Western blotting using anti-ISG15 antibody (1:1,000 dilution). Fold change in band density, as calculated via normalization of ISG15 with β-actin (loading control), was measured using ImageJ software (NIH, Bethesda, MD). (E) ISG15 and USP18 mRNA expression levels in swine liver tissue samples obtained from animals experimentally infected with HEV ( n = 3) at 3 weeks postinfection. The fold changes in swine ISG15 and USP18 mRNA levels were calculated using the 2 −ΔΔ CT method compared to the uninfected control animals ( n = 2), and the RPS18 gene was used as the housekeeping gene. The data represent means ± standard errors of the means (SEM) of results from three independent transfection experiments (A, B, and C), one experiment representative of two independent transfection experiments (D), and HEV-infected animals ( n = 3) compared to uninfected animals ( n = 2) (E). **, P ≤ 0.01.
    Sybr Green Master Mix Thermo Fisher Scientific Cat, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 84/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sybr green master mix thermo fisher scientific cat/product/Thermo Fisher
    Average 84 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    sybr green master mix thermo fisher scientific cat - by Bioz Stars, 2020-07
    84/100 stars
      Buy from Supplier

    99
    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 .
    Sybr Green Master Mix, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 33260 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sybr green master mix/product/Thermo Fisher
    Average 99 stars, based on 33260 article reviews
    Price from $9.99 to $1999.99
    sybr green master mix - by Bioz Stars, 2020-07
    99/100 stars
      Buy from Supplier

    88
    Thermo Fisher mix buffer
    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 .
    Mix Buffer, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 88/100, based on 82 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mix buffer/product/Thermo Fisher
    Average 88 stars, based on 82 article reviews
    Price from $9.99 to $1999.99
    mix buffer - by Bioz Stars, 2020-07
    88/100 stars
      Buy from Supplier

    92
    Bio-Rad 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 .
    Master Mix, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 92/100, based on 258 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/master mix/product/Bio-Rad
    Average 92 stars, based on 258 article reviews
    Price from $9.99 to $1999.99
    master mix - by Bioz Stars, 2020-07
    92/100 stars
      Buy from Supplier

    99
    Thermo Fisher taqman master mix
    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 <t>ISAV</t> standard curve by (A) SYBR GREEN I and <t>(B)TaqMan;</t> Correlation of detection of copy number of ISAV in plasma samples by (C) SYBR Green I (n = 13) and (D) TaqMan chemistries (n = 8).
    Taqman Master Mix, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 4590 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/taqman master mix/product/Thermo Fisher
    Average 99 stars, based on 4590 article reviews
    Price from $9.99 to $1999.99
    taqman master mix - by Bioz Stars, 2020-07
    99/100 stars
      Buy from Supplier

    99
    Thermo Fisher superscript vilo master mix
    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 <t>ISAV</t> standard curve by (A) SYBR GREEN I and <t>(B)TaqMan;</t> Correlation of detection of copy number of ISAV in plasma samples by (C) SYBR Green I (n = 13) and (D) TaqMan chemistries (n = 8).
    Superscript Vilo Master Mix, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 2222 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/superscript vilo master mix/product/Thermo Fisher
    Average 99 stars, based on 2222 article reviews
    Price from $9.99 to $1999.99
    superscript vilo master mix - by Bioz Stars, 2020-07
    99/100 stars
      Buy from Supplier

    90
    Thermo Fisher sybr green mix
    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 <t>ISAV</t> standard curve by (A) SYBR GREEN I and <t>(B)TaqMan;</t> 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 Mix, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 4390 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sybr green mix/product/Thermo Fisher
    Average 90 stars, based on 4390 article reviews
    Price from $9.99 to $1999.99
    sybr green mix - by Bioz Stars, 2020-07
    90/100 stars
      Buy from Supplier

    94
    Thermo Fisher sybrtm select master mix
    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 <t>ISAV</t> standard curve by (A) SYBR GREEN I and <t>(B)TaqMan;</t> Correlation of detection of copy number of ISAV in plasma samples by (C) SYBR Green I (n = 13) and (D) TaqMan chemistries (n = 8).
    Sybrtm Select Master Mix, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 86 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sybrtm select master mix/product/Thermo Fisher
    Average 94 stars, based on 86 article reviews
    Price from $9.99 to $1999.99
    sybrtm select master mix - by Bioz Stars, 2020-07
    94/100 stars
      Buy from Supplier

    Image Search Results


    HEV induces ISG15 both in vitro and in vivo . (A) Replication kinetics of viral RNAs from the HEV P6 infectious cDNA clone and HEV P6GLuc replicon in Huh7-S10-3 liver cells. The cells were transfected with capped RNA transcripts from the genotype 3 HEV P6 infectious cDNA clone (HEV P6) or with the HEV P6GLuc replicon (HEV P6GLuc; P6 encoding Gaussia luciferase clone). The culture supernatants were collected at various time points and used to measure viral RNA replication levels by HEV real-time quantitative RT-PCR or by the Gaussia luciferase assay (GLuc assay). RLU, relative light units. (B) ISG15 mRNA expression levels in Huh7-S10-3 liver cells transfected with capped RNA transcripts from the HEV P6 infectious cDNA clone or the HEV P6GLuc replicon at 1 and 5 days posttransfection (dpt) were determined by Sybr green-based quantitative PCR (qPCR). The ISG15-mRNA fold change was calculated using the 2 −ΔΔ CT method for comparisons to untransfected control cells, and the RPS18 gene was used as the housekeeping gene. (C and D) ISG15 protein expression levels in Huh7-S10-3 cells transfected with capped RNA from the HEV P6 infectious cDNA clone or the HEV P6GLuc replicon (C and D) or the HEV P6 replication-deficient infectious cDNA clone (P6GAD) (D). CC, untransfected cell control. The ISG15 protein expression, at 5 days posttransfection, in cell lysates (40 μg/lane) was analyzed by Western blotting using anti-ISG15 antibody (1:1,000 dilution). Fold change in band density, as calculated via normalization of ISG15 with β-actin (loading control), was measured using ImageJ software (NIH, Bethesda, MD). (E) ISG15 and USP18 mRNA expression levels in swine liver tissue samples obtained from animals experimentally infected with HEV ( n = 3) at 3 weeks postinfection. The fold changes in swine ISG15 and USP18 mRNA levels were calculated using the 2 −ΔΔ CT method compared to the uninfected control animals ( n = 2), and the RPS18 gene was used as the housekeeping gene. The data represent means ± standard errors of the means (SEM) of results from three independent transfection experiments (A, B, and C), one experiment representative of two independent transfection experiments (D), and HEV-infected animals ( n = 3) compared to uninfected animals ( n = 2) (E). **, P ≤ 0.01.

    Journal: Journal of Virology

    Article Title: ISG15 Modulates Type I Interferon Signaling and the Antiviral Response during Hepatitis E Virus Replication

    doi: 10.1128/JVI.00621-17

    Figure Lengend Snippet: HEV induces ISG15 both in vitro and in vivo . (A) Replication kinetics of viral RNAs from the HEV P6 infectious cDNA clone and HEV P6GLuc replicon in Huh7-S10-3 liver cells. The cells were transfected with capped RNA transcripts from the genotype 3 HEV P6 infectious cDNA clone (HEV P6) or with the HEV P6GLuc replicon (HEV P6GLuc; P6 encoding Gaussia luciferase clone). The culture supernatants were collected at various time points and used to measure viral RNA replication levels by HEV real-time quantitative RT-PCR or by the Gaussia luciferase assay (GLuc assay). RLU, relative light units. (B) ISG15 mRNA expression levels in Huh7-S10-3 liver cells transfected with capped RNA transcripts from the HEV P6 infectious cDNA clone or the HEV P6GLuc replicon at 1 and 5 days posttransfection (dpt) were determined by Sybr green-based quantitative PCR (qPCR). The ISG15-mRNA fold change was calculated using the 2 −ΔΔ CT method for comparisons to untransfected control cells, and the RPS18 gene was used as the housekeeping gene. (C and D) ISG15 protein expression levels in Huh7-S10-3 cells transfected with capped RNA from the HEV P6 infectious cDNA clone or the HEV P6GLuc replicon (C and D) or the HEV P6 replication-deficient infectious cDNA clone (P6GAD) (D). CC, untransfected cell control. The ISG15 protein expression, at 5 days posttransfection, in cell lysates (40 μg/lane) was analyzed by Western blotting using anti-ISG15 antibody (1:1,000 dilution). Fold change in band density, as calculated via normalization of ISG15 with β-actin (loading control), was measured using ImageJ software (NIH, Bethesda, MD). (E) ISG15 and USP18 mRNA expression levels in swine liver tissue samples obtained from animals experimentally infected with HEV ( n = 3) at 3 weeks postinfection. The fold changes in swine ISG15 and USP18 mRNA levels were calculated using the 2 −ΔΔ CT method compared to the uninfected control animals ( n = 2), and the RPS18 gene was used as the housekeeping gene. The data represent means ± standard errors of the means (SEM) of results from three independent transfection experiments (A, B, and C), one experiment representative of two independent transfection experiments (D), and HEV-infected animals ( n = 3) compared to uninfected animals ( n = 2) (E). **, P ≤ 0.01.

    Article Snippet: The mRNA levels of ISG15, OAS1, PKR, Mx1, USP18, and RPS18 (housekeeping control) were determined using SYBR master mix (Applied Biosystems, Grand Island, NY) with gene-specific primer sets ( ) and a Bio-Rad IQ5 system.

    Techniques: In Vitro, In Vivo, Transfection, Luciferase, Quantitative RT-PCR, Expressing, SYBR Green Assay, Real-time Polymerase Chain Reaction, Western Blot, Software, Infection

    ISG15 regulates type I IFN signaling. (A) Relative pSTAT1 levels. Huh7-S10-3 liver cells were transfected with 20 nM control siRNA (siCnt) or ISG15-siRNA (siISG15) or UBE1L-siRNA plus UBE2L6-siRNA (siE1E2). At 24 hpt, cells were treated with IFN-α (100 IU/ml) for 30 min to 7 h. Cell lysate (20 μg/lane) was analyzed by Western blotting with the indicated antibody, anti-pSTAT1 (1:1,000 dilution), anti-STAT1 (1:1,000 dilution), and anti-β-actin (1:1,000 dilution). Fold change in band intensity was determined using ImageJ software (NIH, Bethesda, MD). The data represent means ± SEM of results from five independent experiments. a, P ≤ 0.05; aa, P ≤ 0.01 (compared with siCnt plus IFN-α at the given time point). (B) IFN-stimulated response element (ISRE) promoter activity levels. Huh7-S10-3 liver cells were cotransfected with siCnt/siISG15/siE1E2 along with pGL4.45[luc2P/ISRE/Hygro] (firefly luciferase) and pGL4.74[hRluc/TK] (renilla luciferase). At 24 hpt, cells were treated with various concentrations of IFN-α. Relative levels of fold induction of the cell-associated firefly luciferase activity, compared to the corresponding untreated cell control levels, at 18 h post-IFN-α treatment were estimated using a dual-luciferase assay kit and were normalized with renilla luciferase expression levels. The data represent means ± SEM of results from triplicate sample experiments. aa, P ≤ 0.01 (compared to siCnt plus IFN-α). (C to F) ISG mRNA levels in siRNA-transfected and IFN-α-treated samples were measured for Mx1 (C), OAS1 (D), PKR (E), and ISG15 (F) using Sybr green qPCR. Fold change in mRNA levels compared to the untransfected control was calculated using the 2 −ΔΔ CT method, and the RPS18 gene was used as the housekeeping gene. The data represent means ± SEM of results from three independent experiments. a, P ≤ 0.05; aa, P ≤ 0.01; aaa, P ≤ 0.001 (compared to siCnt plus IFN-α).

    Journal: Journal of Virology

    Article Title: ISG15 Modulates Type I Interferon Signaling and the Antiviral Response during Hepatitis E Virus Replication

    doi: 10.1128/JVI.00621-17

    Figure Lengend Snippet: ISG15 regulates type I IFN signaling. (A) Relative pSTAT1 levels. Huh7-S10-3 liver cells were transfected with 20 nM control siRNA (siCnt) or ISG15-siRNA (siISG15) or UBE1L-siRNA plus UBE2L6-siRNA (siE1E2). At 24 hpt, cells were treated with IFN-α (100 IU/ml) for 30 min to 7 h. Cell lysate (20 μg/lane) was analyzed by Western blotting with the indicated antibody, anti-pSTAT1 (1:1,000 dilution), anti-STAT1 (1:1,000 dilution), and anti-β-actin (1:1,000 dilution). Fold change in band intensity was determined using ImageJ software (NIH, Bethesda, MD). The data represent means ± SEM of results from five independent experiments. a, P ≤ 0.05; aa, P ≤ 0.01 (compared with siCnt plus IFN-α at the given time point). (B) IFN-stimulated response element (ISRE) promoter activity levels. Huh7-S10-3 liver cells were cotransfected with siCnt/siISG15/siE1E2 along with pGL4.45[luc2P/ISRE/Hygro] (firefly luciferase) and pGL4.74[hRluc/TK] (renilla luciferase). At 24 hpt, cells were treated with various concentrations of IFN-α. Relative levels of fold induction of the cell-associated firefly luciferase activity, compared to the corresponding untreated cell control levels, at 18 h post-IFN-α treatment were estimated using a dual-luciferase assay kit and were normalized with renilla luciferase expression levels. The data represent means ± SEM of results from triplicate sample experiments. aa, P ≤ 0.01 (compared to siCnt plus IFN-α). (C to F) ISG mRNA levels in siRNA-transfected and IFN-α-treated samples were measured for Mx1 (C), OAS1 (D), PKR (E), and ISG15 (F) using Sybr green qPCR. Fold change in mRNA levels compared to the untransfected control was calculated using the 2 −ΔΔ CT method, and the RPS18 gene was used as the housekeeping gene. The data represent means ± SEM of results from three independent experiments. a, P ≤ 0.05; aa, P ≤ 0.01; aaa, P ≤ 0.001 (compared to siCnt plus IFN-α).

    Article Snippet: The mRNA levels of ISG15, OAS1, PKR, Mx1, USP18, and RPS18 (housekeeping control) were determined using SYBR master mix (Applied Biosystems, Grand Island, NY) with gene-specific primer sets ( ) and a Bio-Rad IQ5 system.

    Techniques: Transfection, Western Blot, Software, Activity Assay, Luciferase, Expressing, SYBR Green Assay, Real-time Polymerase Chain Reaction

    Gene expression analysis of fatty acid-metabolizing enzymes in control and H-RasV12 expressing fibroblasts by qRT-PCR. ( A ) Gene expression analysis of desaturases ( SCD and FADS ), elongases ( ELOVL ) and acyl-coenzyme A synthetases ( ACSL ). ( B ) Gene expression analysis of phospholipases A2 ( PLA2 ). Ten ng of each cDNA were used as template. Reactions were performed in triplicate, using SYBR green technology StepOne RT-PCR machine to detect amplification. The GAPDH gene was used as endogenous control. The fold expression in H-RasV12 fibroblasts with respect to control is displayed, expressed as Relative Quantity (RQ). The analysis was repeated three times in triplicate and the mean ± S.D. is reported (* p

    Journal: International Journal of Molecular Sciences

    Article Title: Oncogenic H-Ras Expression Induces Fatty Acid Profile Changes in Human Fibroblasts and Extracellular Vesicles

    doi: 10.3390/ijms19113515

    Figure Lengend Snippet: Gene expression analysis of fatty acid-metabolizing enzymes in control and H-RasV12 expressing fibroblasts by qRT-PCR. ( A ) Gene expression analysis of desaturases ( SCD and FADS ), elongases ( ELOVL ) and acyl-coenzyme A synthetases ( ACSL ). ( B ) Gene expression analysis of phospholipases A2 ( PLA2 ). Ten ng of each cDNA were used as template. Reactions were performed in triplicate, using SYBR green technology StepOne RT-PCR machine to detect amplification. The GAPDH gene was used as endogenous control. The fold expression in H-RasV12 fibroblasts with respect to control is displayed, expressed as Relative Quantity (RQ). The analysis was repeated three times in triplicate and the mean ± S.D. is reported (* p

    Article Snippet: RNA was extracted and retro-transcribed as previously described [ ]. cDNA was used to determine the expression of genes listed in . cDNA was used to determine transcript levels by qRT-PCR in a StepOne RT-PCR machine (Applied Biosystems, Foster City, CA, USA) using SYBR® Select Master Mix (Life Technologies, Carlsbad, CA, USA).

    Techniques: Expressing, Quantitative RT-PCR, SYBR Green Assay, Step One RT-PCR, Amplification

    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 .

    Journal: PLoS ONE

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

    doi: 10.1371/journal.pone.0035461

    Figure Lengend 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 .

    Article Snippet: Real-time PCR was performed in an ABI Prism 7000 Sequence Detection System using SYBR Green Master Mix (Applied Biosystems).

    Techniques: 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

    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: The PCR mix for the Taqman assay consisted of 5 μl of cDNA dilution and 15 μl of master mix prepared using 1 μl of a 20X TaqMan probe/primer mix (5 μM probe: FAM-labelled ISAV segment 8 and 18 μM each primers) [ ], 10 μl TaqMan master mix (Applied Biosystems) and 4 μl of nuclease free water.

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