taqman detection system Thermo Fisher Search Results


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  • 99
    Thermo Fisher taqman fast virus 1 step master mix
    Comparison of the analytical sensitivity for Methods A (used the <t>TaqMan®</t> Fast Virus 1-Step Master Mix (Applied Biosystems®)) and B (used the Superscript III Platinum® One-Step qRT-PCR Kit (Invitrogen ™ )). C T values are the average of three replicates, and bars represent standard deviation. : Method A, : Method B.
    Taqman Fast Virus 1 Step Master Mix, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1555 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher abi prism 7900ht sequence detection system
    Comparison of the analytical sensitivity for Methods A (used the <t>TaqMan®</t> Fast Virus 1-Step Master Mix (Applied Biosystems®)) and B (used the Superscript III Platinum® One-Step qRT-PCR Kit (Invitrogen ™ )). C T values are the average of three replicates, and bars represent standard deviation. : Method A, : Method B.
    Abi Prism 7900ht Sequence Detection System, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 23368 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher β actin
    PCa cells express EpoR. (A) EpoR mRNA levels of PCa cell lines (PC3 and C4-2B) and RWPE1 cells were determined by real time RT-PCR. Data were normalized to <t>β-actin</t> and are presented as mean ± SEM from three independent PCRs. Representative
    β Actin, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 18155 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher taqman environmental master mix 2 0
    PCa cells express EpoR. (A) EpoR mRNA levels of PCa cell lines (PC3 and C4-2B) and RWPE1 cells were determined by real time RT-PCR. Data were normalized to <t>β-actin</t> and are presented as mean ± SEM from three independent PCRs. Representative
    Taqman Environmental Master Mix 2 0, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 642 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher taqman microrna assay
    Nuclear factor erythroid 2-related factor 2 (Nrf2)-related microRNAs are upregulated in the infarcted heart. Tissues from the noninfarcted area of the left ventricle were collected and subjected to RNA extraction and quantification of mature microRNAs using specific <t>TaqMan</t> <t>microRNA</t> assays, including microRNA-27a, microRNA-28a, and microRNA-34a ( A ). B : quantification of other mature microRNAs, including microRNA-142-5p, microRNA-144, and microRNA-153. U6 snRNA was used as an internal control (means ± SE). CHF, chronic heart failure.
    Taqman Microrna Assay, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 12087 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher taqman gene expression assays
    Knockdown of SRSF3 decreased the expression of miR-1908 and its host gene, FADS1 A . Microarray data showing decreased expression of FADS1 in SRSF3-silenced U2OS cells with three different siRNAs. B . U2OS cells were transfected with control (CTRL) or SRSF3-specific siRNA for 48 h and then the expression level of FADS1 mRNA was determined by RT-qPCR analysis. C-D . The levels of pri-miR-1908 and mature miR-1908-3p/5p were assessed by RT-qPCR analysis using pri-miR-1908-specific primers and <t>TaqMan</t> <t>miRNA</t> primers, respectively. E . Cells were transfected with control (CTRL) or FADS1-specific siRNA for 48 h and then the levels of pri-miR-1908 and mature miR-1908-3p/5p were assessed by RT-qPCR as described above. F . Cells were transfected with control (CTRL) or Sp1-specific siRNA for 48 h. The expression level of FADS1 mRNA was determined by RT-qPCR analysis and the levels of pri-miR-1908 and mature miR-1908 (3p/5p) were assessed by RT-qPCR analysis as described above. All experiments are performed more than three times and data represent the mean ± S.D. Asterisk (*) indicates statistical significance of p
    Taqman Gene Expression Assays, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 48363 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    96
    Thermo Fisher taqman rnase p detection reagents kit
    Knockdown of SRSF3 decreased the expression of miR-1908 and its host gene, FADS1 A . Microarray data showing decreased expression of FADS1 in SRSF3-silenced U2OS cells with three different siRNAs. B . U2OS cells were transfected with control (CTRL) or SRSF3-specific siRNA for 48 h and then the expression level of FADS1 mRNA was determined by RT-qPCR analysis. C-D . The levels of pri-miR-1908 and mature miR-1908-3p/5p were assessed by RT-qPCR analysis using pri-miR-1908-specific primers and <t>TaqMan</t> <t>miRNA</t> primers, respectively. E . Cells were transfected with control (CTRL) or FADS1-specific siRNA for 48 h and then the levels of pri-miR-1908 and mature miR-1908-3p/5p were assessed by RT-qPCR as described above. F . Cells were transfected with control (CTRL) or Sp1-specific siRNA for 48 h. The expression level of FADS1 mRNA was determined by RT-qPCR analysis and the levels of pri-miR-1908 and mature miR-1908 (3p/5p) were assessed by RT-qPCR analysis as described above. All experiments are performed more than three times and data represent the mean ± S.D. Asterisk (*) indicates statistical significance of p
    Taqman Rnase P Detection Reagents Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 96/100, based on 294 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher abi prism 7000 sequence detection system
    Knockdown of SRSF3 decreased the expression of miR-1908 and its host gene, FADS1 A . Microarray data showing decreased expression of FADS1 in SRSF3-silenced U2OS cells with three different siRNAs. B . U2OS cells were transfected with control (CTRL) or SRSF3-specific siRNA for 48 h and then the expression level of FADS1 mRNA was determined by RT-qPCR analysis. C-D . The levels of pri-miR-1908 and mature miR-1908-3p/5p were assessed by RT-qPCR analysis using pri-miR-1908-specific primers and <t>TaqMan</t> <t>miRNA</t> primers, respectively. E . Cells were transfected with control (CTRL) or FADS1-specific siRNA for 48 h and then the levels of pri-miR-1908 and mature miR-1908-3p/5p were assessed by RT-qPCR as described above. F . Cells were transfected with control (CTRL) or Sp1-specific siRNA for 48 h. The expression level of FADS1 mRNA was determined by RT-qPCR analysis and the levels of pri-miR-1908 and mature miR-1908 (3p/5p) were assessed by RT-qPCR analysis as described above. All experiments are performed more than three times and data represent the mean ± S.D. Asterisk (*) indicates statistical significance of p
    Abi Prism 7000 Sequence Detection System, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 26130 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher 7900ht sequence detection system
    Knockdown of SRSF3 decreased the expression of miR-1908 and its host gene, FADS1 A . Microarray data showing decreased expression of FADS1 in SRSF3-silenced U2OS cells with three different siRNAs. B . U2OS cells were transfected with control (CTRL) or SRSF3-specific siRNA for 48 h and then the expression level of FADS1 mRNA was determined by RT-qPCR analysis. C-D . The levels of pri-miR-1908 and mature miR-1908-3p/5p were assessed by RT-qPCR analysis using pri-miR-1908-specific primers and <t>TaqMan</t> <t>miRNA</t> primers, respectively. E . Cells were transfected with control (CTRL) or FADS1-specific siRNA for 48 h and then the levels of pri-miR-1908 and mature miR-1908-3p/5p were assessed by RT-qPCR as described above. F . Cells were transfected with control (CTRL) or Sp1-specific siRNA for 48 h. The expression level of FADS1 mRNA was determined by RT-qPCR analysis and the levels of pri-miR-1908 and mature miR-1908 (3p/5p) were assessed by RT-qPCR analysis as described above. All experiments are performed more than three times and data represent the mean ± S.D. Asterisk (*) indicates statistical significance of p
    7900ht Sequence Detection System, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 6352 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher taqman gmo soy 35s detection kit
    Knockdown of SRSF3 decreased the expression of miR-1908 and its host gene, FADS1 A . Microarray data showing decreased expression of FADS1 in SRSF3-silenced U2OS cells with three different siRNAs. B . U2OS cells were transfected with control (CTRL) or SRSF3-specific siRNA for 48 h and then the expression level of FADS1 mRNA was determined by RT-qPCR analysis. C-D . The levels of pri-miR-1908 and mature miR-1908-3p/5p were assessed by RT-qPCR analysis using pri-miR-1908-specific primers and <t>TaqMan</t> <t>miRNA</t> primers, respectively. E . Cells were transfected with control (CTRL) or FADS1-specific siRNA for 48 h and then the levels of pri-miR-1908 and mature miR-1908-3p/5p were assessed by RT-qPCR as described above. F . Cells were transfected with control (CTRL) or Sp1-specific siRNA for 48 h. The expression level of FADS1 mRNA was determined by RT-qPCR analysis and the levels of pri-miR-1908 and mature miR-1908 (3p/5p) were assessed by RT-qPCR analysis as described above. All experiments are performed more than three times and data represent the mean ± S.D. Asterisk (*) indicates statistical significance of p
    Taqman Gmo Soy 35s Detection Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 84/100, based on 50 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher taqman probes
    Knockdown of SRSF3 decreased the expression of miR-1908 and its host gene, FADS1 A . Microarray data showing decreased expression of FADS1 in SRSF3-silenced U2OS cells with three different siRNAs. B . U2OS cells were transfected with control (CTRL) or SRSF3-specific siRNA for 48 h and then the expression level of FADS1 mRNA was determined by RT-qPCR analysis. C-D . The levels of pri-miR-1908 and mature miR-1908-3p/5p were assessed by RT-qPCR analysis using pri-miR-1908-specific primers and <t>TaqMan</t> <t>miRNA</t> primers, respectively. E . Cells were transfected with control (CTRL) or FADS1-specific siRNA for 48 h and then the levels of pri-miR-1908 and mature miR-1908-3p/5p were assessed by RT-qPCR as described above. F . Cells were transfected with control (CTRL) or Sp1-specific siRNA for 48 h. The expression level of FADS1 mRNA was determined by RT-qPCR analysis and the levels of pri-miR-1908 and mature miR-1908 (3p/5p) were assessed by RT-qPCR analysis as described above. All experiments are performed more than three times and data represent the mean ± S.D. Asterisk (*) indicates statistical significance of p
    Taqman Probes, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 30308 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher taqman universal pcr master mix
    Validation of ifnar1 real-time <t>RT-PCR</t> . Ifnar1 detection assay was based on the use of <t>TaqMan</t> probes. Validation curves were performed with RNA extracted from HeLa cells, and the threshold cycle difference ( ΔCT ) between ifnar1 and the constitutive endogenous gene hprt was calculated and drawn with respect to RNA concentration. Slope value ( y ) indicates that hprt can be used for relative quantification of ifnar1 transcription.
    Taqman Universal Pcr Master Mix, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 54482 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher taqman assays
    EBV miR-BHRF1-2-5p contributes to the growth of EBV+ DLBCL cells. A. <t>Taqman</t> qRT-PCR analysis of miR-BHRF1-2-5p (5p) and miR-BHRF1-2-3p (3p) expression in miR-BHRF1-2-5p sponged DLBCLs (IBL1 and BCKN1). Values are normalized to U6 and reported relative to the BHRF1-2 miRNA levels in each respective DLBCL transduced with pLCE-CXCR4 control vector. B. Proliferation of DLBCLs as determined by MTT assay following stable transduction with pLCE-CXCR4 control vector or the miR-BHRF1-2-5p sponge. FAC-sorted, GFP+ DLBCLs were maintained in media containing 15% FBS and split one day prior to MTT assays. Values at Tn are normalized to the absorbance values at 0 hr (A-T0). Error bars represent S.D. of measurements from six or eight wells. *By Student’s t test, p
    Taqman Assays, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 16795 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher abi prism 7700 sequence detection system
    EBV miR-BHRF1-2-5p contributes to the growth of EBV+ DLBCL cells. A. <t>Taqman</t> qRT-PCR analysis of miR-BHRF1-2-5p (5p) and miR-BHRF1-2-3p (3p) expression in miR-BHRF1-2-5p sponged DLBCLs (IBL1 and BCKN1). Values are normalized to U6 and reported relative to the BHRF1-2 miRNA levels in each respective DLBCL transduced with pLCE-CXCR4 control vector. B. Proliferation of DLBCLs as determined by MTT assay following stable transduction with pLCE-CXCR4 control vector or the miR-BHRF1-2-5p sponge. FAC-sorted, GFP+ DLBCLs were maintained in media containing 15% FBS and split one day prior to MTT assays. Values at Tn are normalized to the absorbance values at 0 hr (A-T0). Error bars represent S.D. of measurements from six or eight wells. *By Student’s t test, p
    Abi Prism 7700 Sequence Detection System, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 18083 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher taqman mutation detection assay
    EBV miR-BHRF1-2-5p contributes to the growth of EBV+ DLBCL cells. A. <t>Taqman</t> qRT-PCR analysis of miR-BHRF1-2-5p (5p) and miR-BHRF1-2-3p (3p) expression in miR-BHRF1-2-5p sponged DLBCLs (IBL1 and BCKN1). Values are normalized to U6 and reported relative to the BHRF1-2 miRNA levels in each respective DLBCL transduced with pLCE-CXCR4 control vector. B. Proliferation of DLBCLs as determined by MTT assay following stable transduction with pLCE-CXCR4 control vector or the miR-BHRF1-2-5p sponge. FAC-sorted, GFP+ DLBCLs were maintained in media containing 15% FBS and split one day prior to MTT assays. Values at Tn are normalized to the absorbance values at 0 hr (A-T0). Error bars represent S.D. of measurements from six or eight wells. *By Student’s t test, p
    Taqman Mutation Detection Assay, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 98/100, based on 76 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher taqman gene expression assay
    EBV miR-BHRF1-2-5p contributes to the growth of EBV+ DLBCL cells. A. <t>Taqman</t> qRT-PCR analysis of miR-BHRF1-2-5p (5p) and miR-BHRF1-2-3p (3p) expression in miR-BHRF1-2-5p sponged DLBCLs (IBL1 and BCKN1). Values are normalized to U6 and reported relative to the BHRF1-2 miRNA levels in each respective DLBCL transduced with pLCE-CXCR4 control vector. B. Proliferation of DLBCLs as determined by MTT assay following stable transduction with pLCE-CXCR4 control vector or the miR-BHRF1-2-5p sponge. FAC-sorted, GFP+ DLBCLs were maintained in media containing 15% FBS and split one day prior to MTT assays. Values at Tn are normalized to the absorbance values at 0 hr (A-T0). Error bars represent S.D. of measurements from six or eight wells. *By Student’s t test, p
    Taqman Gene Expression Assay, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 12710 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Comparison of the analytical sensitivity for Methods A (used the TaqMan® Fast Virus 1-Step Master Mix (Applied Biosystems®)) and B (used the Superscript III Platinum® One-Step qRT-PCR Kit (Invitrogen ™ )). C T values are the average of three replicates, and bars represent standard deviation. : Method A, : Method B.

    Journal: Veterinary Microbiology

    Article Title: Detection of foot-and-mouth disease virus in milk samples by real-time reverse transcription polymerase chain reaction: Optimisation and evaluation of a high-throughput screening method with potential for disease surveillance

    doi: 10.1016/j.vetmic.2018.07.024

    Figure Lengend Snippet: Comparison of the analytical sensitivity for Methods A (used the TaqMan® Fast Virus 1-Step Master Mix (Applied Biosystems®)) and B (used the Superscript III Platinum® One-Step qRT-PCR Kit (Invitrogen ™ )). C T values are the average of three replicates, and bars represent standard deviation. : Method A, : Method B.

    Article Snippet: In Method A, the TaqMan® Fast Virus 1-Step Master Mix (Applied Biosystems®) was used with the following thermal cycling conditions: 50 °C for 5 min, 95 °C for 20 s, then 45 cycles of 95 °C for 3 s and 60 °C for 30 s. For this method 2.5 μL of RNA template was added to the rRT-PCR reaction mix containing 6.25 μL of 1-step mastermix (4x, supplied with the kit), 0.25 μL each of forward and reverse primer (20 μM), 0.25 μL probe (10 μM), and 14.5 μL of nuclease free water.

    Techniques: Quantitative RT-PCR, Standard Deviation

    PCa cells express EpoR. (A) EpoR mRNA levels of PCa cell lines (PC3 and C4-2B) and RWPE1 cells were determined by real time RT-PCR. Data were normalized to β-actin and are presented as mean ± SEM from three independent PCRs. Representative

    Journal: Journal of cellular biochemistry

    Article Title: Erythropoietin Supports the Survival of Prostate Cancer, But Not Growth and Bone Metastasis

    doi: 10.1002/jcb.24592

    Figure Lengend Snippet: PCa cells express EpoR. (A) EpoR mRNA levels of PCa cell lines (PC3 and C4-2B) and RWPE1 cells were determined by real time RT-PCR. Data were normalized to β-actin and are presented as mean ± SEM from three independent PCRs. Representative

    Article Snippet: RT products were analyzed by real-time PCR in TaqMan® Gene Expression Assays for human EpoR and β-actin (Applied Biosystems, Foster City, CA).

    Techniques: Quantitative RT-PCR

    The 5’ UTR mutants are defective for Gag/Pol and Env protein expression. Western blot analysis of HEK293T cells transfected with the wild type (WT) and indicated mutant clones. (A) α- MMTV CA (Blue 7) antibody. (B) Env-specific antibody (α-MTV Env) with human β-actin serving as a loading control in both gels. Mock, HEK293T cells transfected with pcDNA3 plasmid alone.

    Journal: Journal of molecular biology

    Article Title: A cis-acting Element Downstream of the Mouse Mammary Tumor Virus Major Splice Donor Critical for RNA Elongation and Stability

    doi: 10.1016/j.jmb.2018.08.025

    Figure Lengend Snippet: The 5’ UTR mutants are defective for Gag/Pol and Env protein expression. Western blot analysis of HEK293T cells transfected with the wild type (WT) and indicated mutant clones. (A) α- MMTV CA (Blue 7) antibody. (B) Env-specific antibody (α-MTV Env) with human β-actin serving as a loading control in both gels. Mock, HEK293T cells transfected with pcDNA3 plasmid alone.

    Article Snippet: The human β-actin Endogenous Control Assay (VIC/MGB probe, Cat# 4326315E, Applied Biosystems (ABI), USA) was used as the control in assays where cellular cDNA (2 μl/sample) served as the template, whereas the human β-actin Endogenous Control Assay (FAM/MGB probe, Cat# 401846, ABI,) was used as the endogenous control with 50 μg gDNA/sample as the template.

    Techniques: Expressing, Western Blot, Transfection, Mutagenesis, Plasmid Preparation

    Nuclear factor erythroid 2-related factor 2 (Nrf2)-related microRNAs are upregulated in the infarcted heart. Tissues from the noninfarcted area of the left ventricle were collected and subjected to RNA extraction and quantification of mature microRNAs using specific TaqMan microRNA assays, including microRNA-27a, microRNA-28a, and microRNA-34a ( A ). B : quantification of other mature microRNAs, including microRNA-142-5p, microRNA-144, and microRNA-153. U6 snRNA was used as an internal control (means ± SE). CHF, chronic heart failure.

    Journal: American Journal of Physiology - Heart and Circulatory Physiology

    Article Title: Signaling and Stress Response: Myocardial infarction-induced microRNA-enriched exosomes contribute to cardiac Nrf2 dysregulation in chronic heart failure

    doi: 10.1152/ajpheart.00602.2017

    Figure Lengend Snippet: Nuclear factor erythroid 2-related factor 2 (Nrf2)-related microRNAs are upregulated in the infarcted heart. Tissues from the noninfarcted area of the left ventricle were collected and subjected to RNA extraction and quantification of mature microRNAs using specific TaqMan microRNA assays, including microRNA-27a, microRNA-28a, and microRNA-34a ( A ). B : quantification of other mature microRNAs, including microRNA-142-5p, microRNA-144, and microRNA-153. U6 snRNA was used as an internal control (means ± SE). CHF, chronic heart failure.

    Article Snippet: MicroRNA-27a, microRNA-28-3p, microRNA-34a, cel-microRNA-39, and U6 snRNA were detected using Taqman microRNA assays (Applied Biosystems) on a StepOne Real-Time PCR systems (Applied Biosystems).

    Techniques: RNA Extraction

    nuclear factor erythroid 2-related factor 2 (Nrf2)-related microRNAs exhibit no significant changes in extra cardiac tissues. Tissues from the kidney, skeletal muscle, and whole brain stem were collected and subjected to RNA extraction and quantification of mature microRNAs using specific TaqMan microRNA assays, including microRNA-27a ( A , D , and G ), microRNA-28-3p ( B , E , and H ), and microRNA-34a ( C , F , and I ). U6 snRNA was used as an internal control (means ± SE). CHF, chronic heart failure.

    Journal: American Journal of Physiology - Heart and Circulatory Physiology

    Article Title: Signaling and Stress Response: Myocardial infarction-induced microRNA-enriched exosomes contribute to cardiac Nrf2 dysregulation in chronic heart failure

    doi: 10.1152/ajpheart.00602.2017

    Figure Lengend Snippet: nuclear factor erythroid 2-related factor 2 (Nrf2)-related microRNAs exhibit no significant changes in extra cardiac tissues. Tissues from the kidney, skeletal muscle, and whole brain stem were collected and subjected to RNA extraction and quantification of mature microRNAs using specific TaqMan microRNA assays, including microRNA-27a ( A , D , and G ), microRNA-28-3p ( B , E , and H ), and microRNA-34a ( C , F , and I ). U6 snRNA was used as an internal control (means ± SE). CHF, chronic heart failure.

    Article Snippet: MicroRNA-27a, microRNA-28-3p, microRNA-34a, cel-microRNA-39, and U6 snRNA were detected using Taqman microRNA assays (Applied Biosystems) on a StepOne Real-Time PCR systems (Applied Biosystems).

    Techniques: RNA Extraction

    Knockdown of SRSF3 decreased the expression of miR-1908 and its host gene, FADS1 A . Microarray data showing decreased expression of FADS1 in SRSF3-silenced U2OS cells with three different siRNAs. B . U2OS cells were transfected with control (CTRL) or SRSF3-specific siRNA for 48 h and then the expression level of FADS1 mRNA was determined by RT-qPCR analysis. C-D . The levels of pri-miR-1908 and mature miR-1908-3p/5p were assessed by RT-qPCR analysis using pri-miR-1908-specific primers and TaqMan miRNA primers, respectively. E . Cells were transfected with control (CTRL) or FADS1-specific siRNA for 48 h and then the levels of pri-miR-1908 and mature miR-1908-3p/5p were assessed by RT-qPCR as described above. F . Cells were transfected with control (CTRL) or Sp1-specific siRNA for 48 h. The expression level of FADS1 mRNA was determined by RT-qPCR analysis and the levels of pri-miR-1908 and mature miR-1908 (3p/5p) were assessed by RT-qPCR analysis as described above. All experiments are performed more than three times and data represent the mean ± S.D. Asterisk (*) indicates statistical significance of p

    Journal: Oncotarget

    Article Title: MicroRNA-1908-5p contributes to the oncogenic function of the splicing factor SRSF3

    doi: 10.18632/oncotarget.14184

    Figure Lengend Snippet: Knockdown of SRSF3 decreased the expression of miR-1908 and its host gene, FADS1 A . Microarray data showing decreased expression of FADS1 in SRSF3-silenced U2OS cells with three different siRNAs. B . U2OS cells were transfected with control (CTRL) or SRSF3-specific siRNA for 48 h and then the expression level of FADS1 mRNA was determined by RT-qPCR analysis. C-D . The levels of pri-miR-1908 and mature miR-1908-3p/5p were assessed by RT-qPCR analysis using pri-miR-1908-specific primers and TaqMan miRNA primers, respectively. E . Cells were transfected with control (CTRL) or FADS1-specific siRNA for 48 h and then the levels of pri-miR-1908 and mature miR-1908-3p/5p were assessed by RT-qPCR as described above. F . Cells were transfected with control (CTRL) or Sp1-specific siRNA for 48 h. The expression level of FADS1 mRNA was determined by RT-qPCR analysis and the levels of pri-miR-1908 and mature miR-1908 (3p/5p) were assessed by RT-qPCR analysis as described above. All experiments are performed more than three times and data represent the mean ± S.D. Asterisk (*) indicates statistical significance of p

    Article Snippet: For detection of miRNA, TaqMan® gene expression assays were performed using hsa-1908-5p-specific and has-miR-1908-3p specific primers (Applied Biosystems, USA).

    Techniques: Expressing, Microarray, Transfection, Quantitative RT-PCR

    NF-κB is involved in SRSF3-regulated miR-1908 expression A . U2OS cells were transfected with control (CTRL) or SRSF3-specific siRNA. Transfected cells were resuspended into 12-well plates and then transfected with the NF-κB reporter vector. Transactivation of NF-κB was assessed by calculating the luciferase activity. B . To check whether NF-κB regulates the expression of miR-1908, cells were treated with the NF-κB-specific inhibitor BAY11-7082 (10 μM) for 24 h. The level of miR-1908-3p/5p was determined by RT-qPCR analysis using TaqMan primers. C . Cells were transfected with control (CTRL) or SRSF3-specific siRNA for 48 h and then whole cell lysates were prepared. Western blot analyses were performed using adequate specific antibodies. D . Cells were transfected as described above and then cellular fractionation was performed. Western blot analysis was performed to determine the localization of NF-κB (p65) and α-tubulin and lamin B was checked for cytoplasmic and nuclear markers, respectively. E-F . Cells were transfected with control (CTRL) or TAK1-specific siRNA and then the levels of TAK1 and phosphorylated IKKα/β and NF-κB were assessed by Western blot analysis. The level of miR-1908-5p was determined by RT-qPCR analysis. G-H . The protein and mRNA expression levels of NF-κB target genes including HIF1-α, cyclin D1, and slug, were determined by western blot and RT-qPCR analysis, respectively. All experiments are performed more than three times and data represent the mean ± S.D. Asterisk (*) indicates statistical significance of p

    Journal: Oncotarget

    Article Title: MicroRNA-1908-5p contributes to the oncogenic function of the splicing factor SRSF3

    doi: 10.18632/oncotarget.14184

    Figure Lengend Snippet: NF-κB is involved in SRSF3-regulated miR-1908 expression A . U2OS cells were transfected with control (CTRL) or SRSF3-specific siRNA. Transfected cells were resuspended into 12-well plates and then transfected with the NF-κB reporter vector. Transactivation of NF-κB was assessed by calculating the luciferase activity. B . To check whether NF-κB regulates the expression of miR-1908, cells were treated with the NF-κB-specific inhibitor BAY11-7082 (10 μM) for 24 h. The level of miR-1908-3p/5p was determined by RT-qPCR analysis using TaqMan primers. C . Cells were transfected with control (CTRL) or SRSF3-specific siRNA for 48 h and then whole cell lysates were prepared. Western blot analyses were performed using adequate specific antibodies. D . Cells were transfected as described above and then cellular fractionation was performed. Western blot analysis was performed to determine the localization of NF-κB (p65) and α-tubulin and lamin B was checked for cytoplasmic and nuclear markers, respectively. E-F . Cells were transfected with control (CTRL) or TAK1-specific siRNA and then the levels of TAK1 and phosphorylated IKKα/β and NF-κB were assessed by Western blot analysis. The level of miR-1908-5p was determined by RT-qPCR analysis. G-H . The protein and mRNA expression levels of NF-κB target genes including HIF1-α, cyclin D1, and slug, were determined by western blot and RT-qPCR analysis, respectively. All experiments are performed more than three times and data represent the mean ± S.D. Asterisk (*) indicates statistical significance of p

    Article Snippet: For detection of miRNA, TaqMan® gene expression assays were performed using hsa-1908-5p-specific and has-miR-1908-3p specific primers (Applied Biosystems, USA).

    Techniques: Expressing, Transfection, Plasmid Preparation, Luciferase, Activity Assay, Quantitative RT-PCR, Western Blot, Cell Fractionation

    Validation of ifnar1 real-time RT-PCR . Ifnar1 detection assay was based on the use of TaqMan probes. Validation curves were performed with RNA extracted from HeLa cells, and the threshold cycle difference ( ΔCT ) between ifnar1 and the constitutive endogenous gene hprt was calculated and drawn with respect to RNA concentration. Slope value ( y ) indicates that hprt can be used for relative quantification of ifnar1 transcription.

    Journal: Virology Journal

    Article Title: Quantitative analysis of interferon alpha receptor subunit 1 and suppressor of cytokine signaling 1 gene transcription in blood cells of patients with chronic hepatitis C

    doi: 10.1186/1743-422X-7-243

    Figure Lengend Snippet: Validation of ifnar1 real-time RT-PCR . Ifnar1 detection assay was based on the use of TaqMan probes. Validation curves were performed with RNA extracted from HeLa cells, and the threshold cycle difference ( ΔCT ) between ifnar1 and the constitutive endogenous gene hprt was calculated and drawn with respect to RNA concentration. Slope value ( y ) indicates that hprt can be used for relative quantification of ifnar1 transcription.

    Article Snippet: Real-time RT-PCR for ifnar1 All RT-PCR were set up in 96-well optical plates using 4 ng of patients' RNA, 10 μl TaqMan Universal PCR Master Mix (Applied Biosystems, Foster City, CA, USA), and 1 μl of primers/probe set containing 900 nM of forward (5'-GCTTTGGATGGTTTAAGCTTTACATATAGC-3'), and reverse primers (5'-TCTGGTGAGAGTTTATAAATTTTATGTCTGGAAT-3'), and 300 nM probe [FAM]CTTCAGGTGTAGAAGAAAG[NFQ] was added to a final volume of 20 μl per reaction.

    Techniques: Quantitative RT-PCR, Detection Assay, Concentration Assay

    EBV miR-BHRF1-2-5p contributes to the growth of EBV+ DLBCL cells. A. Taqman qRT-PCR analysis of miR-BHRF1-2-5p (5p) and miR-BHRF1-2-3p (3p) expression in miR-BHRF1-2-5p sponged DLBCLs (IBL1 and BCKN1). Values are normalized to U6 and reported relative to the BHRF1-2 miRNA levels in each respective DLBCL transduced with pLCE-CXCR4 control vector. B. Proliferation of DLBCLs as determined by MTT assay following stable transduction with pLCE-CXCR4 control vector or the miR-BHRF1-2-5p sponge. FAC-sorted, GFP+ DLBCLs were maintained in media containing 15% FBS and split one day prior to MTT assays. Values at Tn are normalized to the absorbance values at 0 hr (A-T0). Error bars represent S.D. of measurements from six or eight wells. *By Student’s t test, p

    Journal: PLoS Pathogens

    Article Title: Epstein-Barr virus microRNAs regulate B cell receptor signal transduction and lytic reactivation

    doi: 10.1371/journal.ppat.1007535

    Figure Lengend Snippet: EBV miR-BHRF1-2-5p contributes to the growth of EBV+ DLBCL cells. A. Taqman qRT-PCR analysis of miR-BHRF1-2-5p (5p) and miR-BHRF1-2-3p (3p) expression in miR-BHRF1-2-5p sponged DLBCLs (IBL1 and BCKN1). Values are normalized to U6 and reported relative to the BHRF1-2 miRNA levels in each respective DLBCL transduced with pLCE-CXCR4 control vector. B. Proliferation of DLBCLs as determined by MTT assay following stable transduction with pLCE-CXCR4 control vector or the miR-BHRF1-2-5p sponge. FAC-sorted, GFP+ DLBCLs were maintained in media containing 15% FBS and split one day prior to MTT assays. Values at Tn are normalized to the absorbance values at 0 hr (A-T0). Error bars represent S.D. of measurements from six or eight wells. *By Student’s t test, p

    Article Snippet: Oligonucleotides are listed in Table S1. miRNAs were detected using commercial Taqman assays or miRNA stem-loop qRT-PCR assays as previously described [ , ]. miRNA levels are reported relative to miR-16 (assay #000391, Thermofisher) or U6 (assay #001973, Thermofisher) as indicated.

    Techniques: Quantitative RT-PCR, Expressing, Transduction, Plasmid Preparation, MTT Assay