taqman detection system Thermo Fisher Search Results


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  • 90
    Thermo Fisher taqman mutation detection assays
    Representative results for <t>EGFR</t> mutation screening using direct sequencing, fragment analysis, real-time allelic discrimination, Therascreen EGFR RGQ kit, and EGFR <t>TaqMan</t> Mutation Detection Assays. (a) The right panel is an example of wild type EGFR, the left panel is an example of L858R mutation. (b) The right panel is an example of wild type EGFR, the left panel is an example of a deletion in exon 19 (c.2237_2254del18bp).
    Taqman Mutation Detection Assays, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 40 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher taqman detection system
    Representative results for <t>EGFR</t> mutation screening using direct sequencing, fragment analysis, real-time allelic discrimination, Therascreen EGFR RGQ kit, and EGFR <t>TaqMan</t> Mutation Detection Assays. (a) The right panel is an example of wild type EGFR, the left panel is an example of L858R mutation. (b) The right panel is an example of wild type EGFR, the left panel is an example of a deletion in exon 19 (c.2237_2254del18bp).
    Taqman Detection System, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 96/100, based on 2793 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher kaiso specific taqman
    <t>Kaiso</t> is required for EGF-induced cell migration and invasion. A: shRNA Kaiso down-regulated Kaiso levels at mRNA as determined by quantitative RT-PCR with Kaiso-specific <t>TaqMan</t> primers and HPRT1 as the loading control or protein levels by immunoblot.
    Kaiso Specific Taqman, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 75/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher taqman based detection chemistries
    <t>Kaiso</t> is required for EGF-induced cell migration and invasion. A: shRNA Kaiso down-regulated Kaiso levels at mRNA as determined by quantitative RT-PCR with Kaiso-specific <t>TaqMan</t> primers and HPRT1 as the loading control or protein levels by immunoblot.
    Taqman Based Detection Chemistries, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 78/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher taqman gmo maize 35s detection kit
    <t>Kaiso</t> is required for EGF-induced cell migration and invasion. A: shRNA Kaiso down-regulated Kaiso levels at mRNA as determined by quantitative RT-PCR with Kaiso-specific <t>TaqMan</t> primers and HPRT1 as the loading control or protein levels by immunoblot.
    Taqman Gmo Maize 35s Detection Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 77/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher taqman detection probes
    <t>Kaiso</t> is required for EGF-induced cell migration and invasion. A: shRNA Kaiso down-regulated Kaiso levels at mRNA as determined by quantitative RT-PCR with Kaiso-specific <t>TaqMan</t> primers and HPRT1 as the loading control or protein levels by immunoblot.
    Taqman Detection Probes, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 77/100, based on 799 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher fluorescent taqman probe
    <t>Kaiso</t> is required for EGF-induced cell migration and invasion. A: shRNA Kaiso down-regulated Kaiso levels at mRNA as determined by quantitative RT-PCR with Kaiso-specific <t>TaqMan</t> primers and HPRT1 as the loading control or protein levels by immunoblot.
    Fluorescent Taqman Probe, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 82/100, based on 33 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher sybr green taqman assay
    <t>Kaiso</t> is required for EGF-induced cell migration and invasion. A: shRNA Kaiso down-regulated Kaiso levels at mRNA as determined by quantitative RT-PCR with Kaiso-specific <t>TaqMan</t> primers and HPRT1 as the loading control or protein levels by immunoblot.
    Sybr Green Taqman Assay, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 86/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher taqman detection systems
    <t>Kaiso</t> is required for EGF-induced cell migration and invasion. A: shRNA Kaiso down-regulated Kaiso levels at mRNA as determined by quantitative RT-PCR with Kaiso-specific <t>TaqMan</t> primers and HPRT1 as the loading control or protein levels by immunoblot.
    Taqman Detection Systems, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 87/100, based on 31 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher taqman assay by design service
    <t>Kaiso</t> is required for EGF-induced cell migration and invasion. A: shRNA Kaiso down-regulated Kaiso levels at mRNA as determined by quantitative RT-PCR with Kaiso-specific <t>TaqMan</t> primers and HPRT1 as the loading control or protein levels by immunoblot.
    Taqman Assay By Design Service, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 81/100, based on 22 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher taqman snp assay
    Correlation of XCI patterns obtained with the use of 3 different methods . (A) HUMARA assay versus <t>TaqMan</t> <t>SNP</t> assay at the IDS locus. For each individual, the %Sup measured by HUMARA was compared with the percentage of IDS allele obtained with the TaqMan
    Taqman Snp Assay, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 206 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher taqman fluorogenic detection system
    Co-culture of encapsulated WT vs KO murine preadipocytes alter gene expression in canine adipocytes Schematic presentation of an experiment in which encapsulated murine preadipocytes were co-cultured and differentiated with adherent canine preadipocytes (A). The upper panel shows encapsulated murine preadipocytes floating in media under light microscopy (20x magnification). In schematics, differentiated canine adipocytes were depicted as adherent cells containing yellow lipid droplets. Numerous murine adipocytes (blue ellipses) are floating in the media within microcapsules (circles) Canine preadipocytes (Mastiff dog) were differentiated in adipogenic medium with encapsulated WT or Aldh1a1 −/− (KO) preadipocytes for 14 days (B-F). Expression of Pparg (B), Fabp4 (C), Atgl (D), Ucp1 (E) and LOC479623 (F) were measured by <t>Q-PCR</t> assay <t>TaqMan.</t> The values were normalized by TBP and represent the mean ± SD (n=3), one-way ANOVA.
    Taqman Fluorogenic Detection System, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 62 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher taqman hydrolysis probes
    Co-culture of encapsulated WT vs KO murine preadipocytes alter gene expression in canine adipocytes Schematic presentation of an experiment in which encapsulated murine preadipocytes were co-cultured and differentiated with adherent canine preadipocytes (A). The upper panel shows encapsulated murine preadipocytes floating in media under light microscopy (20x magnification). In schematics, differentiated canine adipocytes were depicted as adherent cells containing yellow lipid droplets. Numerous murine adipocytes (blue ellipses) are floating in the media within microcapsules (circles) Canine preadipocytes (Mastiff dog) were differentiated in adipogenic medium with encapsulated WT or Aldh1a1 −/− (KO) preadipocytes for 14 days (B-F). Expression of Pparg (B), Fabp4 (C), Atgl (D), Ucp1 (E) and LOC479623 (F) were measured by <t>Q-PCR</t> assay <t>TaqMan.</t> The values were normalized by TBP and represent the mean ± SD (n=3), one-way ANOVA.
    Taqman Hydrolysis Probes, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 98/100, based on 173 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher mir 9 taqman
    Co-culture of encapsulated WT vs KO murine preadipocytes alter gene expression in canine adipocytes Schematic presentation of an experiment in which encapsulated murine preadipocytes were co-cultured and differentiated with adherent canine preadipocytes (A). The upper panel shows encapsulated murine preadipocytes floating in media under light microscopy (20x magnification). In schematics, differentiated canine adipocytes were depicted as adherent cells containing yellow lipid droplets. Numerous murine adipocytes (blue ellipses) are floating in the media within microcapsules (circles) Canine preadipocytes (Mastiff dog) were differentiated in adipogenic medium with encapsulated WT or Aldh1a1 −/− (KO) preadipocytes for 14 days (B-F). Expression of Pparg (B), Fabp4 (C), Atgl (D), Ucp1 (E) and LOC479623 (F) were measured by <t>Q-PCR</t> assay <t>TaqMan.</t> The values were normalized by TBP and represent the mean ± SD (n=3), one-way ANOVA.
    Mir 9 Taqman, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 77/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher taqman microrna assay
    Co-culture of encapsulated WT vs KO murine preadipocytes alter gene expression in canine adipocytes Schematic presentation of an experiment in which encapsulated murine preadipocytes were co-cultured and differentiated with adherent canine preadipocytes (A). The upper panel shows encapsulated murine preadipocytes floating in media under light microscopy (20x magnification). In schematics, differentiated canine adipocytes were depicted as adherent cells containing yellow lipid droplets. Numerous murine adipocytes (blue ellipses) are floating in the media within microcapsules (circles) Canine preadipocytes (Mastiff dog) were differentiated in adipogenic medium with encapsulated WT or Aldh1a1 −/− (KO) preadipocytes for 14 days (B-F). Expression of Pparg (B), Fabp4 (C), Atgl (D), Ucp1 (E) and LOC479623 (F) were measured by <t>Q-PCR</t> assay <t>TaqMan.</t> The values were normalized by TBP and represent the mean ± SD (n=3), one-way ANOVA.
    Taqman Microrna Assay, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 4100 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher taqman openarray system
    Co-culture of encapsulated WT vs KO murine preadipocytes alter gene expression in canine adipocytes Schematic presentation of an experiment in which encapsulated murine preadipocytes were co-cultured and differentiated with adherent canine preadipocytes (A). The upper panel shows encapsulated murine preadipocytes floating in media under light microscopy (20x magnification). In schematics, differentiated canine adipocytes were depicted as adherent cells containing yellow lipid droplets. Numerous murine adipocytes (blue ellipses) are floating in the media within microcapsules (circles) Canine preadipocytes (Mastiff dog) were differentiated in adipogenic medium with encapsulated WT or Aldh1a1 −/− (KO) preadipocytes for 14 days (B-F). Expression of Pparg (B), Fabp4 (C), Atgl (D), Ucp1 (E) and LOC479623 (F) were measured by <t>Q-PCR</t> assay <t>TaqMan.</t> The values were normalized by TBP and represent the mean ± SD (n=3), one-way ANOVA.
    Taqman Openarray System, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 41 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher taqman salmonella enterica detection kit
    Co-culture of encapsulated WT vs KO murine preadipocytes alter gene expression in canine adipocytes Schematic presentation of an experiment in which encapsulated murine preadipocytes were co-cultured and differentiated with adherent canine preadipocytes (A). The upper panel shows encapsulated murine preadipocytes floating in media under light microscopy (20x magnification). In schematics, differentiated canine adipocytes were depicted as adherent cells containing yellow lipid droplets. Numerous murine adipocytes (blue ellipses) are floating in the media within microcapsules (circles) Canine preadipocytes (Mastiff dog) were differentiated in adipogenic medium with encapsulated WT or Aldh1a1 −/− (KO) preadipocytes for 14 days (B-F). Expression of Pparg (B), Fabp4 (C), Atgl (D), Ucp1 (E) and LOC479623 (F) were measured by <t>Q-PCR</t> assay <t>TaqMan.</t> The values were normalized by TBP and represent the mean ± SD (n=3), one-way ANOVA.
    Taqman Salmonella Enterica Detection Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 80/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher taqman staphylococcus aureus detection kit
    Detecting <t>RNase</t> P-specific target by PCR. Comparison of phenol-chloroform ( A ), spin column ( B ), and Automate Express ( C ) methods for purifying nucleic acids from blood samples spiked with different quantities of SPS and the use of this material in <t>TaqMan</t> assays for the detection of human RNase P. Samples were processed in triplicate. Data are expressed as means ± SD. IPC, internal positive control; SPS, sodium polyanetholsulfonate.
    Taqman Staphylococcus Aureus Detection Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 79/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher taqman microrna systems
    Differential expression level of miR-142-3p in fibroblasts and iPS. (a) Expression of miR-142-3p was examined by RT-qPCR in various cells. Total RNA was extracted from indicated cells, and RT-qPCR was done using <t>TaqMan</t> <t>MicroRNA</t> systems. U6 shRNA was used as a control. Experiments were done three times using independently prepared cells, and average values with standard deviation are shown. (b) Schematic representation of antisense- (as-) miR-142-3p and EGFP expression plasmid. LTR was used to drive EGFP, and U6 promoter was used to drive as-miR-142-3p. (c) Effect of overexpressed as-miR-142-3p for expression level of endogenous miR-142-3p in iPS cells. as-miR-142-3p/EGFP or control vector was transfected into iPS, and, after 24 hours, level of miR-142-3p in iPS was examined by RT-qPCR. Data were expressed as relative expression level of miR-142-3p in as-miR-142-3p/EGFP expressing cells to that in control vector expressing cells. Experiments were performed three times, and average values with standard deviation are shown. (d, e, and f) Effects of expression of as-miR-142-3p for proliferation and alkaline phosphatase (ALP) expression of iPS. as-miR-142-3p/EGFP plasmid was transfected into undifferentiated iPS, and EGFP positive cells were purified by a cell sorter. Then EGFP positive cells were cultured for 2 days for Ki67 immunostaining and for 5 days for ALP assay. Immunostaining with anti-Ki67 antibody or ALP staining was done, and positive cells were counted under a microscope. Experiments were performed three times, and average values with standard deviation are shown. In (f), morphology of representative colonies of as-miR-142-3p or control vector transfected iPS is shown. (g–i) Expression of lineage marker genes in embryoid body (EB). iPS cells were transfected with as-miR-142-3p/EGFP or as-miR-17/EGFP as a control, purified according to their expression of EGFP, and then subjected to an EB formation. After 6 days of culturing in EB formation condition, the differentiation of cells into the ectodermal (g), endodermal (h), and mesodermal (i) lineages was assessed using RT-qPCR with primers against Fgf5 , Gata4 , and T brachyury , respectively. P value, ∗
    Taqman Microrna Systems, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher taqman 7700 sequencer detection system
    Differential expression level of miR-142-3p in fibroblasts and iPS. (a) Expression of miR-142-3p was examined by RT-qPCR in various cells. Total RNA was extracted from indicated cells, and RT-qPCR was done using <t>TaqMan</t> <t>MicroRNA</t> systems. U6 shRNA was used as a control. Experiments were done three times using independently prepared cells, and average values with standard deviation are shown. (b) Schematic representation of antisense- (as-) miR-142-3p and EGFP expression plasmid. LTR was used to drive EGFP, and U6 promoter was used to drive as-miR-142-3p. (c) Effect of overexpressed as-miR-142-3p for expression level of endogenous miR-142-3p in iPS cells. as-miR-142-3p/EGFP or control vector was transfected into iPS, and, after 24 hours, level of miR-142-3p in iPS was examined by RT-qPCR. Data were expressed as relative expression level of miR-142-3p in as-miR-142-3p/EGFP expressing cells to that in control vector expressing cells. Experiments were performed three times, and average values with standard deviation are shown. (d, e, and f) Effects of expression of as-miR-142-3p for proliferation and alkaline phosphatase (ALP) expression of iPS. as-miR-142-3p/EGFP plasmid was transfected into undifferentiated iPS, and EGFP positive cells were purified by a cell sorter. Then EGFP positive cells were cultured for 2 days for Ki67 immunostaining and for 5 days for ALP assay. Immunostaining with anti-Ki67 antibody or ALP staining was done, and positive cells were counted under a microscope. Experiments were performed three times, and average values with standard deviation are shown. In (f), morphology of representative colonies of as-miR-142-3p or control vector transfected iPS is shown. (g–i) Expression of lineage marker genes in embryoid body (EB). iPS cells were transfected with as-miR-142-3p/EGFP or as-miR-17/EGFP as a control, purified according to their expression of EGFP, and then subjected to an EB formation. After 6 days of culturing in EB formation condition, the differentiation of cells into the ectodermal (g), endodermal (h), and mesodermal (i) lineages was assessed using RT-qPCR with primers against Fgf5 , Gata4 , and T brachyury , respectively. P value, ∗
    Taqman 7700 Sequencer Detection System, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 78/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher taqman mature mir assays
    Differential expression level of miR-142-3p in fibroblasts and iPS. (a) Expression of miR-142-3p was examined by RT-qPCR in various cells. Total RNA was extracted from indicated cells, and RT-qPCR was done using <t>TaqMan</t> <t>MicroRNA</t> systems. U6 shRNA was used as a control. Experiments were done three times using independently prepared cells, and average values with standard deviation are shown. (b) Schematic representation of antisense- (as-) miR-142-3p and EGFP expression plasmid. LTR was used to drive EGFP, and U6 promoter was used to drive as-miR-142-3p. (c) Effect of overexpressed as-miR-142-3p for expression level of endogenous miR-142-3p in iPS cells. as-miR-142-3p/EGFP or control vector was transfected into iPS, and, after 24 hours, level of miR-142-3p in iPS was examined by RT-qPCR. Data were expressed as relative expression level of miR-142-3p in as-miR-142-3p/EGFP expressing cells to that in control vector expressing cells. Experiments were performed three times, and average values with standard deviation are shown. (d, e, and f) Effects of expression of as-miR-142-3p for proliferation and alkaline phosphatase (ALP) expression of iPS. as-miR-142-3p/EGFP plasmid was transfected into undifferentiated iPS, and EGFP positive cells were purified by a cell sorter. Then EGFP positive cells were cultured for 2 days for Ki67 immunostaining and for 5 days for ALP assay. Immunostaining with anti-Ki67 antibody or ALP staining was done, and positive cells were counted under a microscope. Experiments were performed three times, and average values with standard deviation are shown. In (f), morphology of representative colonies of as-miR-142-3p or control vector transfected iPS is shown. (g–i) Expression of lineage marker genes in embryoid body (EB). iPS cells were transfected with as-miR-142-3p/EGFP or as-miR-17/EGFP as a control, purified according to their expression of EGFP, and then subjected to an EB formation. After 6 days of culturing in EB formation condition, the differentiation of cells into the ectodermal (g), endodermal (h), and mesodermal (i) lineages was assessed using RT-qPCR with primers against Fgf5 , Gata4 , and T brachyury , respectively. P value, ∗
    Taqman Mature Mir Assays, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 87/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher taqman based detection primers
    Differential expression level of miR-142-3p in fibroblasts and iPS. (a) Expression of miR-142-3p was examined by RT-qPCR in various cells. Total RNA was extracted from indicated cells, and RT-qPCR was done using <t>TaqMan</t> <t>MicroRNA</t> systems. U6 shRNA was used as a control. Experiments were done three times using independently prepared cells, and average values with standard deviation are shown. (b) Schematic representation of antisense- (as-) miR-142-3p and EGFP expression plasmid. LTR was used to drive EGFP, and U6 promoter was used to drive as-miR-142-3p. (c) Effect of overexpressed as-miR-142-3p for expression level of endogenous miR-142-3p in iPS cells. as-miR-142-3p/EGFP or control vector was transfected into iPS, and, after 24 hours, level of miR-142-3p in iPS was examined by RT-qPCR. Data were expressed as relative expression level of miR-142-3p in as-miR-142-3p/EGFP expressing cells to that in control vector expressing cells. Experiments were performed three times, and average values with standard deviation are shown. (d, e, and f) Effects of expression of as-miR-142-3p for proliferation and alkaline phosphatase (ALP) expression of iPS. as-miR-142-3p/EGFP plasmid was transfected into undifferentiated iPS, and EGFP positive cells were purified by a cell sorter. Then EGFP positive cells were cultured for 2 days for Ki67 immunostaining and for 5 days for ALP assay. Immunostaining with anti-Ki67 antibody or ALP staining was done, and positive cells were counted under a microscope. Experiments were performed three times, and average values with standard deviation are shown. In (f), morphology of representative colonies of as-miR-142-3p or control vector transfected iPS is shown. (g–i) Expression of lineage marker genes in embryoid body (EB). iPS cells were transfected with as-miR-142-3p/EGFP or as-miR-17/EGFP as a control, purified according to their expression of EGFP, and then subjected to an EB formation. After 6 days of culturing in EB formation condition, the differentiation of cells into the ectodermal (g), endodermal (h), and mesodermal (i) lineages was assessed using RT-qPCR with primers against Fgf5 , Gata4 , and T brachyury , respectively. P value, ∗
    Taqman Based Detection Primers, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 79/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher taqman 7900ht detection system
    Differential expression level of miR-142-3p in fibroblasts and iPS. (a) Expression of miR-142-3p was examined by RT-qPCR in various cells. Total RNA was extracted from indicated cells, and RT-qPCR was done using <t>TaqMan</t> <t>MicroRNA</t> systems. U6 shRNA was used as a control. Experiments were done three times using independently prepared cells, and average values with standard deviation are shown. (b) Schematic representation of antisense- (as-) miR-142-3p and EGFP expression plasmid. LTR was used to drive EGFP, and U6 promoter was used to drive as-miR-142-3p. (c) Effect of overexpressed as-miR-142-3p for expression level of endogenous miR-142-3p in iPS cells. as-miR-142-3p/EGFP or control vector was transfected into iPS, and, after 24 hours, level of miR-142-3p in iPS was examined by RT-qPCR. Data were expressed as relative expression level of miR-142-3p in as-miR-142-3p/EGFP expressing cells to that in control vector expressing cells. Experiments were performed three times, and average values with standard deviation are shown. (d, e, and f) Effects of expression of as-miR-142-3p for proliferation and alkaline phosphatase (ALP) expression of iPS. as-miR-142-3p/EGFP plasmid was transfected into undifferentiated iPS, and EGFP positive cells were purified by a cell sorter. Then EGFP positive cells were cultured for 2 days for Ki67 immunostaining and for 5 days for ALP assay. Immunostaining with anti-Ki67 antibody or ALP staining was done, and positive cells were counted under a microscope. Experiments were performed three times, and average values with standard deviation are shown. In (f), morphology of representative colonies of as-miR-142-3p or control vector transfected iPS is shown. (g–i) Expression of lineage marker genes in embryoid body (EB). iPS cells were transfected with as-miR-142-3p/EGFP or as-miR-17/EGFP as a control, purified according to their expression of EGFP, and then subjected to an EB formation. After 6 days of culturing in EB formation condition, the differentiation of cells into the ectodermal (g), endodermal (h), and mesodermal (i) lineages was assessed using RT-qPCR with primers against Fgf5 , Gata4 , and T brachyury , respectively. P value, ∗
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    Differential expression level of miR-142-3p in fibroblasts and iPS. (a) Expression of miR-142-3p was examined by RT-qPCR in various cells. Total RNA was extracted from indicated cells, and RT-qPCR was done using <t>TaqMan</t> <t>MicroRNA</t> systems. U6 shRNA was used as a control. Experiments were done three times using independently prepared cells, and average values with standard deviation are shown. (b) Schematic representation of antisense- (as-) miR-142-3p and EGFP expression plasmid. LTR was used to drive EGFP, and U6 promoter was used to drive as-miR-142-3p. (c) Effect of overexpressed as-miR-142-3p for expression level of endogenous miR-142-3p in iPS cells. as-miR-142-3p/EGFP or control vector was transfected into iPS, and, after 24 hours, level of miR-142-3p in iPS was examined by RT-qPCR. Data were expressed as relative expression level of miR-142-3p in as-miR-142-3p/EGFP expressing cells to that in control vector expressing cells. Experiments were performed three times, and average values with standard deviation are shown. (d, e, and f) Effects of expression of as-miR-142-3p for proliferation and alkaline phosphatase (ALP) expression of iPS. as-miR-142-3p/EGFP plasmid was transfected into undifferentiated iPS, and EGFP positive cells were purified by a cell sorter. Then EGFP positive cells were cultured for 2 days for Ki67 immunostaining and for 5 days for ALP assay. Immunostaining with anti-Ki67 antibody or ALP staining was done, and positive cells were counted under a microscope. Experiments were performed three times, and average values with standard deviation are shown. In (f), morphology of representative colonies of as-miR-142-3p or control vector transfected iPS is shown. (g–i) Expression of lineage marker genes in embryoid body (EB). iPS cells were transfected with as-miR-142-3p/EGFP or as-miR-17/EGFP as a control, purified according to their expression of EGFP, and then subjected to an EB formation. After 6 days of culturing in EB formation condition, the differentiation of cells into the ectodermal (g), endodermal (h), and mesodermal (i) lineages was assessed using RT-qPCR with primers against Fgf5 , Gata4 , and T brachyury , respectively. P value, ∗
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    Differential expression level of miR-142-3p in fibroblasts and iPS. (a) Expression of miR-142-3p was examined by RT-qPCR in various cells. Total RNA was extracted from indicated cells, and RT-qPCR was done using <t>TaqMan</t> <t>MicroRNA</t> systems. U6 shRNA was used as a control. Experiments were done three times using independently prepared cells, and average values with standard deviation are shown. (b) Schematic representation of antisense- (as-) miR-142-3p and EGFP expression plasmid. LTR was used to drive EGFP, and U6 promoter was used to drive as-miR-142-3p. (c) Effect of overexpressed as-miR-142-3p for expression level of endogenous miR-142-3p in iPS cells. as-miR-142-3p/EGFP or control vector was transfected into iPS, and, after 24 hours, level of miR-142-3p in iPS was examined by RT-qPCR. Data were expressed as relative expression level of miR-142-3p in as-miR-142-3p/EGFP expressing cells to that in control vector expressing cells. Experiments were performed three times, and average values with standard deviation are shown. (d, e, and f) Effects of expression of as-miR-142-3p for proliferation and alkaline phosphatase (ALP) expression of iPS. as-miR-142-3p/EGFP plasmid was transfected into undifferentiated iPS, and EGFP positive cells were purified by a cell sorter. Then EGFP positive cells were cultured for 2 days for Ki67 immunostaining and for 5 days for ALP assay. Immunostaining with anti-Ki67 antibody or ALP staining was done, and positive cells were counted under a microscope. Experiments were performed three times, and average values with standard deviation are shown. In (f), morphology of representative colonies of as-miR-142-3p or control vector transfected iPS is shown. (g–i) Expression of lineage marker genes in embryoid body (EB). iPS cells were transfected with as-miR-142-3p/EGFP or as-miR-17/EGFP as a control, purified according to their expression of EGFP, and then subjected to an EB formation. After 6 days of culturing in EB formation condition, the differentiation of cells into the ectodermal (g), endodermal (h), and mesodermal (i) lineages was assessed using RT-qPCR with primers against Fgf5 , Gata4 , and T brachyury , respectively. P value, ∗
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    Differential expression level of miR-142-3p in fibroblasts and iPS. (a) Expression of miR-142-3p was examined by RT-qPCR in various cells. Total RNA was extracted from indicated cells, and RT-qPCR was done using <t>TaqMan</t> <t>MicroRNA</t> systems. U6 shRNA was used as a control. Experiments were done three times using independently prepared cells, and average values with standard deviation are shown. (b) Schematic representation of antisense- (as-) miR-142-3p and EGFP expression plasmid. LTR was used to drive EGFP, and U6 promoter was used to drive as-miR-142-3p. (c) Effect of overexpressed as-miR-142-3p for expression level of endogenous miR-142-3p in iPS cells. as-miR-142-3p/EGFP or control vector was transfected into iPS, and, after 24 hours, level of miR-142-3p in iPS was examined by RT-qPCR. Data were expressed as relative expression level of miR-142-3p in as-miR-142-3p/EGFP expressing cells to that in control vector expressing cells. Experiments were performed three times, and average values with standard deviation are shown. (d, e, and f) Effects of expression of as-miR-142-3p for proliferation and alkaline phosphatase (ALP) expression of iPS. as-miR-142-3p/EGFP plasmid was transfected into undifferentiated iPS, and EGFP positive cells were purified by a cell sorter. Then EGFP positive cells were cultured for 2 days for Ki67 immunostaining and for 5 days for ALP assay. Immunostaining with anti-Ki67 antibody or ALP staining was done, and positive cells were counted under a microscope. Experiments were performed three times, and average values with standard deviation are shown. In (f), morphology of representative colonies of as-miR-142-3p or control vector transfected iPS is shown. (g–i) Expression of lineage marker genes in embryoid body (EB). iPS cells were transfected with as-miR-142-3p/EGFP or as-miR-17/EGFP as a control, purified according to their expression of EGFP, and then subjected to an EB formation. After 6 days of culturing in EB formation condition, the differentiation of cells into the ectodermal (g), endodermal (h), and mesodermal (i) lineages was assessed using RT-qPCR with primers against Fgf5 , Gata4 , and T brachyury , respectively. P value, ∗
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    Differential expression level of miR-142-3p in fibroblasts and iPS. (a) Expression of miR-142-3p was examined by RT-qPCR in various cells. Total RNA was extracted from indicated cells, and RT-qPCR was done using <t>TaqMan</t> <t>MicroRNA</t> systems. U6 shRNA was used as a control. Experiments were done three times using independently prepared cells, and average values with standard deviation are shown. (b) Schematic representation of antisense- (as-) miR-142-3p and EGFP expression plasmid. LTR was used to drive EGFP, and U6 promoter was used to drive as-miR-142-3p. (c) Effect of overexpressed as-miR-142-3p for expression level of endogenous miR-142-3p in iPS cells. as-miR-142-3p/EGFP or control vector was transfected into iPS, and, after 24 hours, level of miR-142-3p in iPS was examined by RT-qPCR. Data were expressed as relative expression level of miR-142-3p in as-miR-142-3p/EGFP expressing cells to that in control vector expressing cells. Experiments were performed three times, and average values with standard deviation are shown. (d, e, and f) Effects of expression of as-miR-142-3p for proliferation and alkaline phosphatase (ALP) expression of iPS. as-miR-142-3p/EGFP plasmid was transfected into undifferentiated iPS, and EGFP positive cells were purified by a cell sorter. Then EGFP positive cells were cultured for 2 days for Ki67 immunostaining and for 5 days for ALP assay. Immunostaining with anti-Ki67 antibody or ALP staining was done, and positive cells were counted under a microscope. Experiments were performed three times, and average values with standard deviation are shown. In (f), morphology of representative colonies of as-miR-142-3p or control vector transfected iPS is shown. (g–i) Expression of lineage marker genes in embryoid body (EB). iPS cells were transfected with as-miR-142-3p/EGFP or as-miR-17/EGFP as a control, purified according to their expression of EGFP, and then subjected to an EB formation. After 6 days of culturing in EB formation condition, the differentiation of cells into the ectodermal (g), endodermal (h), and mesodermal (i) lineages was assessed using RT-qPCR with primers against Fgf5 , Gata4 , and T brachyury , respectively. P value, ∗
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    Differential expression level of miR-142-3p in fibroblasts and iPS. (a) Expression of miR-142-3p was examined by RT-qPCR in various cells. Total RNA was extracted from indicated cells, and RT-qPCR was done using <t>TaqMan</t> <t>MicroRNA</t> systems. U6 shRNA was used as a control. Experiments were done three times using independently prepared cells, and average values with standard deviation are shown. (b) Schematic representation of antisense- (as-) miR-142-3p and EGFP expression plasmid. LTR was used to drive EGFP, and U6 promoter was used to drive as-miR-142-3p. (c) Effect of overexpressed as-miR-142-3p for expression level of endogenous miR-142-3p in iPS cells. as-miR-142-3p/EGFP or control vector was transfected into iPS, and, after 24 hours, level of miR-142-3p in iPS was examined by RT-qPCR. Data were expressed as relative expression level of miR-142-3p in as-miR-142-3p/EGFP expressing cells to that in control vector expressing cells. Experiments were performed three times, and average values with standard deviation are shown. (d, e, and f) Effects of expression of as-miR-142-3p for proliferation and alkaline phosphatase (ALP) expression of iPS. as-miR-142-3p/EGFP plasmid was transfected into undifferentiated iPS, and EGFP positive cells were purified by a cell sorter. Then EGFP positive cells were cultured for 2 days for Ki67 immunostaining and for 5 days for ALP assay. Immunostaining with anti-Ki67 antibody or ALP staining was done, and positive cells were counted under a microscope. Experiments were performed three times, and average values with standard deviation are shown. In (f), morphology of representative colonies of as-miR-142-3p or control vector transfected iPS is shown. (g–i) Expression of lineage marker genes in embryoid body (EB). iPS cells were transfected with as-miR-142-3p/EGFP or as-miR-17/EGFP as a control, purified according to their expression of EGFP, and then subjected to an EB formation. After 6 days of culturing in EB formation condition, the differentiation of cells into the ectodermal (g), endodermal (h), and mesodermal (i) lineages was assessed using RT-qPCR with primers against Fgf5 , Gata4 , and T brachyury , respectively. P value, ∗
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    Differential expression level of miR-142-3p in fibroblasts and iPS. (a) Expression of miR-142-3p was examined by RT-qPCR in various cells. Total RNA was extracted from indicated cells, and RT-qPCR was done using <t>TaqMan</t> <t>MicroRNA</t> systems. U6 shRNA was used as a control. Experiments were done three times using independently prepared cells, and average values with standard deviation are shown. (b) Schematic representation of antisense- (as-) miR-142-3p and EGFP expression plasmid. LTR was used to drive EGFP, and U6 promoter was used to drive as-miR-142-3p. (c) Effect of overexpressed as-miR-142-3p for expression level of endogenous miR-142-3p in iPS cells. as-miR-142-3p/EGFP or control vector was transfected into iPS, and, after 24 hours, level of miR-142-3p in iPS was examined by RT-qPCR. Data were expressed as relative expression level of miR-142-3p in as-miR-142-3p/EGFP expressing cells to that in control vector expressing cells. Experiments were performed three times, and average values with standard deviation are shown. (d, e, and f) Effects of expression of as-miR-142-3p for proliferation and alkaline phosphatase (ALP) expression of iPS. as-miR-142-3p/EGFP plasmid was transfected into undifferentiated iPS, and EGFP positive cells were purified by a cell sorter. Then EGFP positive cells were cultured for 2 days for Ki67 immunostaining and for 5 days for ALP assay. Immunostaining with anti-Ki67 antibody or ALP staining was done, and positive cells were counted under a microscope. Experiments were performed three times, and average values with standard deviation are shown. In (f), morphology of representative colonies of as-miR-142-3p or control vector transfected iPS is shown. (g–i) Expression of lineage marker genes in embryoid body (EB). iPS cells were transfected with as-miR-142-3p/EGFP or as-miR-17/EGFP as a control, purified according to their expression of EGFP, and then subjected to an EB formation. After 6 days of culturing in EB formation condition, the differentiation of cells into the ectodermal (g), endodermal (h), and mesodermal (i) lineages was assessed using RT-qPCR with primers against Fgf5 , Gata4 , and T brachyury , respectively. P value, ∗
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    Molecular analysis of the patient’s FIP1L1-PDGFRA fusion gene. A: Maps of FIP1L1 and PDGFRA genes. Boxes represent exons according to the LeukemiaNet exons numbering ( FIP1L1 , Ensembl Gene ID: ENSG00000145216; PDGFRA , ENSG00000134853). Numbers below the boxes indicate exon sizes in bp. The previously described FIP1L1 breakpoint region is indicated, encompassing introns 8 to 13, whereas PDGFRA breakpoint is invariably located in exon 12. The white arrow indicates our patient’s FIP1L1 intron 16-peculiar breakpoint. B: Patient’s specific FIP1L1-PDGFRA fusion gene juxtaposing a short portion of FIP1L1 intron 16 (33 bp) to PDGFRA exon 12. Primers and probes used in our assay are indicated. The diagnosis qualitative <t>RT-PCR</t> was performed with the primer pairs FIP1L1-F4 and PDGFRA-R1 for the first round and FIP1L1-F5 combined with PDGFRA-R2 for the second round (Diagnostic primers). Patient’s specific primers used for the qualitative follow-up were FIP1L1S15 and PDGFRAAS13 (Patient’s specific primers). For the quantitative RQ RT-PCR, exon 16 forward primer FIP1L1E16T was used in conjunction with a carboxyfluorescein dye-labeled <t>TaqMan</t> probe, P-PDGF12, and a reverse primer, AS-PDGF12 (RQ primers/probe). C: One of the patient’s specific FIP1L1-PDGFRA sequences represented at the mRNA and protein levels. The fusion gene conserved an open reading frame.
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    Thermo Fisher taqman abi7900ht sequence detector
    Molecular analysis of the patient’s FIP1L1-PDGFRA fusion gene. A: Maps of FIP1L1 and PDGFRA genes. Boxes represent exons according to the LeukemiaNet exons numbering ( FIP1L1 , Ensembl Gene ID: ENSG00000145216; PDGFRA , ENSG00000134853). Numbers below the boxes indicate exon sizes in bp. The previously described FIP1L1 breakpoint region is indicated, encompassing introns 8 to 13, whereas PDGFRA breakpoint is invariably located in exon 12. The white arrow indicates our patient’s FIP1L1 intron 16-peculiar breakpoint. B: Patient’s specific FIP1L1-PDGFRA fusion gene juxtaposing a short portion of FIP1L1 intron 16 (33 bp) to PDGFRA exon 12. Primers and probes used in our assay are indicated. The diagnosis qualitative <t>RT-PCR</t> was performed with the primer pairs FIP1L1-F4 and PDGFRA-R1 for the first round and FIP1L1-F5 combined with PDGFRA-R2 for the second round (Diagnostic primers). Patient’s specific primers used for the qualitative follow-up were FIP1L1S15 and PDGFRAAS13 (Patient’s specific primers). For the quantitative RQ RT-PCR, exon 16 forward primer FIP1L1E16T was used in conjunction with a carboxyfluorescein dye-labeled <t>TaqMan</t> probe, P-PDGF12, and a reverse primer, AS-PDGF12 (RQ primers/probe). C: One of the patient’s specific FIP1L1-PDGFRA sequences represented at the mRNA and protein levels. The fusion gene conserved an open reading frame.
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    Image Search Results


    Representative results for EGFR mutation screening using direct sequencing, fragment analysis, real-time allelic discrimination, Therascreen EGFR RGQ kit, and EGFR TaqMan Mutation Detection Assays. (a) The right panel is an example of wild type EGFR, the left panel is an example of L858R mutation. (b) The right panel is an example of wild type EGFR, the left panel is an example of a deletion in exon 19 (c.2237_2254del18bp).

    Journal: BioMed Research International

    Article Title: Detection of EGFR Mutations by TaqMan Mutation Detection Assays Powered by Competitive Allele-Specific TaqMan PCR Technology

    doi: 10.1155/2013/385087

    Figure Lengend Snippet: Representative results for EGFR mutation screening using direct sequencing, fragment analysis, real-time allelic discrimination, Therascreen EGFR RGQ kit, and EGFR TaqMan Mutation Detection Assays. (a) The right panel is an example of wild type EGFR, the left panel is an example of L858R mutation. (b) The right panel is an example of wild type EGFR, the left panel is an example of a deletion in exon 19 (c.2237_2254del18bp).

    Article Snippet: CastPCR CastPCR was performed in 96-well plates preloaded with TaqMan Mutation Detection Assays, TaqMan EGFR Exon 19 Deletions Assay, and TaqMan Mutation Detection Reference Assays (Life Technologies) in 20 μ L reaction volume including 1x TaqMan genotyping master mix (Life Technologies), deionised water, and 10 ng DNA template.

    Techniques: Mutagenesis, Sequencing

    Kaiso is required for EGF-induced cell migration and invasion. A: shRNA Kaiso down-regulated Kaiso levels at mRNA as determined by quantitative RT-PCR with Kaiso-specific TaqMan primers and HPRT1 as the loading control or protein levels by immunoblot.

    Journal: The American Journal of Pathology

    Article Title: Nuclear Kaiso Indicates Aggressive Prostate Cancers and Promotes Migration and Invasiveness of Prostate Cancer Cells

    doi: 10.1016/j.ajpath.2012.08.008

    Figure Lengend Snippet: Kaiso is required for EGF-induced cell migration and invasion. A: shRNA Kaiso down-regulated Kaiso levels at mRNA as determined by quantitative RT-PCR with Kaiso-specific TaqMan primers and HPRT1 as the loading control or protein levels by immunoblot.

    Article Snippet: RNA was extracted from prostate cancer cells using TRIzol (Invitrogen). cDNA was prepared using Superscript III First Strand cDNA Synthesis kits (Invitrogen) and detected by Kaiso-specific TaqMan (Invitrogen).

    Techniques: Migration, shRNA, Quantitative RT-PCR

    Correlation of XCI patterns obtained with the use of 3 different methods . (A) HUMARA assay versus TaqMan SNP assay at the IDS locus. For each individual, the %Sup measured by HUMARA was compared with the percentage of IDS allele obtained with the TaqMan

    Journal: Blood

    Article Title: Skewing of X-inactivation ratios in blood cells of aging women is confirmed by independent methodologies

    doi: 10.1182/blood-2008-12-195677

    Figure Lengend Snippet: Correlation of XCI patterns obtained with the use of 3 different methods . (A) HUMARA assay versus TaqMan SNP assay at the IDS locus. For each individual, the %Sup measured by HUMARA was compared with the percentage of IDS allele obtained with the TaqMan

    Article Snippet: For this assay, cDNA was synthesized from PMN RNA with the use of random hexamers and TaqMan Reverse Transcription Reagents (Applied Biosystems), after which the aforementioned SNP was detected with the use of the TaqMan SNP assay (Applied Biosystems).

    Techniques:

    Co-culture of encapsulated WT vs KO murine preadipocytes alter gene expression in canine adipocytes Schematic presentation of an experiment in which encapsulated murine preadipocytes were co-cultured and differentiated with adherent canine preadipocytes (A). The upper panel shows encapsulated murine preadipocytes floating in media under light microscopy (20x magnification). In schematics, differentiated canine adipocytes were depicted as adherent cells containing yellow lipid droplets. Numerous murine adipocytes (blue ellipses) are floating in the media within microcapsules (circles) Canine preadipocytes (Mastiff dog) were differentiated in adipogenic medium with encapsulated WT or Aldh1a1 −/− (KO) preadipocytes for 14 days (B-F). Expression of Pparg (B), Fabp4 (C), Atgl (D), Ucp1 (E) and LOC479623 (F) were measured by Q-PCR assay TaqMan. The values were normalized by TBP and represent the mean ± SD (n=3), one-way ANOVA.

    Journal: Xenotransplantation

    Article Title: Aldehyde dehydrogenase 1a1 (Aldh1a1) regulates energy metabolism in adipocytes from different species

    doi: 10.1111/xen.12318

    Figure Lengend Snippet: Co-culture of encapsulated WT vs KO murine preadipocytes alter gene expression in canine adipocytes Schematic presentation of an experiment in which encapsulated murine preadipocytes were co-cultured and differentiated with adherent canine preadipocytes (A). The upper panel shows encapsulated murine preadipocytes floating in media under light microscopy (20x magnification). In schematics, differentiated canine adipocytes were depicted as adherent cells containing yellow lipid droplets. Numerous murine adipocytes (blue ellipses) are floating in the media within microcapsules (circles) Canine preadipocytes (Mastiff dog) were differentiated in adipogenic medium with encapsulated WT or Aldh1a1 −/− (KO) preadipocytes for 14 days (B-F). Expression of Pparg (B), Fabp4 (C), Atgl (D), Ucp1 (E) and LOC479623 (F) were measured by Q-PCR assay TaqMan. The values were normalized by TBP and represent the mean ± SD (n=3), one-way ANOVA.

    Article Snippet: RNA integrity was analyzed using the Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA). mRNA was quantified using 7900HT Fast Real-Time PCR System and TaqMan fluorogenic detection system (Applied Biosystems/ThermoFisher).

    Techniques: Co-Culture Assay, Expressing, Cell Culture, Light Microscopy, Polymerase Chain Reaction

    Co-culture of encapsulated murine WT and KO preadipocytes influence expression of genes in canine adipocytes isolated from a German Shepherd dog Canine preadipocytes were differentiated in adipogenic medium with encapsulated WT or KO preadipocytes for 14 days (A-E). Expression of Pparg (A), Fabp4 (B), Atgl (C), Ucp1 (D) and LOC479623 (E) were measured by Q-PCR assay TaqMan. The values were normalized by TBP and represent the mean ± SD (n=3), one-way ANOVA.

    Journal: Xenotransplantation

    Article Title: Aldehyde dehydrogenase 1a1 (Aldh1a1) regulates energy metabolism in adipocytes from different species

    doi: 10.1111/xen.12318

    Figure Lengend Snippet: Co-culture of encapsulated murine WT and KO preadipocytes influence expression of genes in canine adipocytes isolated from a German Shepherd dog Canine preadipocytes were differentiated in adipogenic medium with encapsulated WT or KO preadipocytes for 14 days (A-E). Expression of Pparg (A), Fabp4 (B), Atgl (C), Ucp1 (D) and LOC479623 (E) were measured by Q-PCR assay TaqMan. The values were normalized by TBP and represent the mean ± SD (n=3), one-way ANOVA.

    Article Snippet: RNA integrity was analyzed using the Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA). mRNA was quantified using 7900HT Fast Real-Time PCR System and TaqMan fluorogenic detection system (Applied Biosystems/ThermoFisher).

    Techniques: Co-Culture Assay, Expressing, Isolation, Polymerase Chain Reaction

    Canine adipocytes express higher levels of differentiation markers PPARg and Fabp4 , and reduced levels of Ucp1 than non-differentiated ASC ASC were isolated from falciform adipose depot of a female Mastiff dog and cultured for 3 passages. The resulted primary non-differentiated pre-adipocytes (ND) were differentiated in adipogenic medium (DMI and DM II) with or without 1µM BRL for 14 days (D for Differentiated). RNA expression of Pparg (A), Fabp4 (B), Atgl (C), Ucp1 (D) and LOC479623 (E) were measured in non-differentiated and differentiated canine adipocytes using Q-PCR assay and TaqMan probes. The values were normalized by TBP and represent the mean ± SD (n=3), one-way ANOVA.

    Journal: Xenotransplantation

    Article Title: Aldehyde dehydrogenase 1a1 (Aldh1a1) regulates energy metabolism in adipocytes from different species

    doi: 10.1111/xen.12318

    Figure Lengend Snippet: Canine adipocytes express higher levels of differentiation markers PPARg and Fabp4 , and reduced levels of Ucp1 than non-differentiated ASC ASC were isolated from falciform adipose depot of a female Mastiff dog and cultured for 3 passages. The resulted primary non-differentiated pre-adipocytes (ND) were differentiated in adipogenic medium (DMI and DM II) with or without 1µM BRL for 14 days (D for Differentiated). RNA expression of Pparg (A), Fabp4 (B), Atgl (C), Ucp1 (D) and LOC479623 (E) were measured in non-differentiated and differentiated canine adipocytes using Q-PCR assay and TaqMan probes. The values were normalized by TBP and represent the mean ± SD (n=3), one-way ANOVA.

    Article Snippet: RNA integrity was analyzed using the Agilent 2100 Bioanalyzer (Agilent Technologies, Santa Clara, CA). mRNA was quantified using 7900HT Fast Real-Time PCR System and TaqMan fluorogenic detection system (Applied Biosystems/ThermoFisher).

    Techniques: Isolation, Cell Culture, RNA Expression, Polymerase Chain Reaction

    Detecting RNase P-specific target by PCR. Comparison of phenol-chloroform ( A ), spin column ( B ), and Automate Express ( C ) methods for purifying nucleic acids from blood samples spiked with different quantities of SPS and the use of this material in TaqMan assays for the detection of human RNase P. Samples were processed in triplicate. Data are expressed as means ± SD. IPC, internal positive control; SPS, sodium polyanetholsulfonate.

    Journal: The Journal of Molecular Diagnostics : JMD

    Article Title: A Sample Extraction Method for Faster, More Sensitive PCR-Based Detection of Pathogens in Blood Culture

    doi: 10.1016/j.jmoldx.2011.10.001

    Figure Lengend Snippet: Detecting RNase P-specific target by PCR. Comparison of phenol-chloroform ( A ), spin column ( B ), and Automate Express ( C ) methods for purifying nucleic acids from blood samples spiked with different quantities of SPS and the use of this material in TaqMan assays for the detection of human RNase P. Samples were processed in triplicate. Data are expressed as means ± SD. IPC, internal positive control; SPS, sodium polyanetholsulfonate.

    Article Snippet: Nucleic acid extracts were screened using TaqMan assays, including a Quantifiler Duo DNA quantification kit (4387746; Applied Biosystems), which measures the levels of ribonuclease P RNA component H1 (RNase P), a TaqMan S. aureus detection kit (4368606; Applied Biosystems), which targets the rnpB gene, and an assay described by Francois et al targeting the mecA gene, which confers resistance to methicillin and its derivatives.

    Techniques: Polymerase Chain Reaction, Positive Control

    Differential expression level of miR-142-3p in fibroblasts and iPS. (a) Expression of miR-142-3p was examined by RT-qPCR in various cells. Total RNA was extracted from indicated cells, and RT-qPCR was done using TaqMan MicroRNA systems. U6 shRNA was used as a control. Experiments were done three times using independently prepared cells, and average values with standard deviation are shown. (b) Schematic representation of antisense- (as-) miR-142-3p and EGFP expression plasmid. LTR was used to drive EGFP, and U6 promoter was used to drive as-miR-142-3p. (c) Effect of overexpressed as-miR-142-3p for expression level of endogenous miR-142-3p in iPS cells. as-miR-142-3p/EGFP or control vector was transfected into iPS, and, after 24 hours, level of miR-142-3p in iPS was examined by RT-qPCR. Data were expressed as relative expression level of miR-142-3p in as-miR-142-3p/EGFP expressing cells to that in control vector expressing cells. Experiments were performed three times, and average values with standard deviation are shown. (d, e, and f) Effects of expression of as-miR-142-3p for proliferation and alkaline phosphatase (ALP) expression of iPS. as-miR-142-3p/EGFP plasmid was transfected into undifferentiated iPS, and EGFP positive cells were purified by a cell sorter. Then EGFP positive cells were cultured for 2 days for Ki67 immunostaining and for 5 days for ALP assay. Immunostaining with anti-Ki67 antibody or ALP staining was done, and positive cells were counted under a microscope. Experiments were performed three times, and average values with standard deviation are shown. In (f), morphology of representative colonies of as-miR-142-3p or control vector transfected iPS is shown. (g–i) Expression of lineage marker genes in embryoid body (EB). iPS cells were transfected with as-miR-142-3p/EGFP or as-miR-17/EGFP as a control, purified according to their expression of EGFP, and then subjected to an EB formation. After 6 days of culturing in EB formation condition, the differentiation of cells into the ectodermal (g), endodermal (h), and mesodermal (i) lineages was assessed using RT-qPCR with primers against Fgf5 , Gata4 , and T brachyury , respectively. P value, ∗

    Journal: Stem Cells International

    Article Title: DNA Methylation Is Involved in the Expression of miR-142-3p in Fibroblasts and Induced Pluripotent Stem Cells

    doi: 10.1155/2014/101349

    Figure Lengend Snippet: Differential expression level of miR-142-3p in fibroblasts and iPS. (a) Expression of miR-142-3p was examined by RT-qPCR in various cells. Total RNA was extracted from indicated cells, and RT-qPCR was done using TaqMan MicroRNA systems. U6 shRNA was used as a control. Experiments were done three times using independently prepared cells, and average values with standard deviation are shown. (b) Schematic representation of antisense- (as-) miR-142-3p and EGFP expression plasmid. LTR was used to drive EGFP, and U6 promoter was used to drive as-miR-142-3p. (c) Effect of overexpressed as-miR-142-3p for expression level of endogenous miR-142-3p in iPS cells. as-miR-142-3p/EGFP or control vector was transfected into iPS, and, after 24 hours, level of miR-142-3p in iPS was examined by RT-qPCR. Data were expressed as relative expression level of miR-142-3p in as-miR-142-3p/EGFP expressing cells to that in control vector expressing cells. Experiments were performed three times, and average values with standard deviation are shown. (d, e, and f) Effects of expression of as-miR-142-3p for proliferation and alkaline phosphatase (ALP) expression of iPS. as-miR-142-3p/EGFP plasmid was transfected into undifferentiated iPS, and EGFP positive cells were purified by a cell sorter. Then EGFP positive cells were cultured for 2 days for Ki67 immunostaining and for 5 days for ALP assay. Immunostaining with anti-Ki67 antibody or ALP staining was done, and positive cells were counted under a microscope. Experiments were performed three times, and average values with standard deviation are shown. In (f), morphology of representative colonies of as-miR-142-3p or control vector transfected iPS is shown. (g–i) Expression of lineage marker genes in embryoid body (EB). iPS cells were transfected with as-miR-142-3p/EGFP or as-miR-17/EGFP as a control, purified according to their expression of EGFP, and then subjected to an EB formation. After 6 days of culturing in EB formation condition, the differentiation of cells into the ectodermal (g), endodermal (h), and mesodermal (i) lineages was assessed using RT-qPCR with primers against Fgf5 , Gata4 , and T brachyury , respectively. P value, ∗

    Article Snippet: RNA Extraction and Real-Time PCR for Quantification of miRNAs and mRNA Total RNA was extracted using the Sepasol (Nacalai Tesque), and level of mature miRNAs was detected using TaqMan MicroRNA systems (Applied Biosystems) using primer specific for each mature miRNA supplied by Applied Biosystems using Light Cycler 1.5 (ROCHE).

    Techniques: Expressing, Quantitative RT-PCR, shRNA, Standard Deviation, Plasmid Preparation, Transfection, ALP Assay, Purification, Cell Culture, Immunostaining, Staining, Microscopy, Marker

    Molecular analysis of the patient’s FIP1L1-PDGFRA fusion gene. A: Maps of FIP1L1 and PDGFRA genes. Boxes represent exons according to the LeukemiaNet exons numbering ( FIP1L1 , Ensembl Gene ID: ENSG00000145216; PDGFRA , ENSG00000134853). Numbers below the boxes indicate exon sizes in bp. The previously described FIP1L1 breakpoint region is indicated, encompassing introns 8 to 13, whereas PDGFRA breakpoint is invariably located in exon 12. The white arrow indicates our patient’s FIP1L1 intron 16-peculiar breakpoint. B: Patient’s specific FIP1L1-PDGFRA fusion gene juxtaposing a short portion of FIP1L1 intron 16 (33 bp) to PDGFRA exon 12. Primers and probes used in our assay are indicated. The diagnosis qualitative RT-PCR was performed with the primer pairs FIP1L1-F4 and PDGFRA-R1 for the first round and FIP1L1-F5 combined with PDGFRA-R2 for the second round (Diagnostic primers). Patient’s specific primers used for the qualitative follow-up were FIP1L1S15 and PDGFRAAS13 (Patient’s specific primers). For the quantitative RQ RT-PCR, exon 16 forward primer FIP1L1E16T was used in conjunction with a carboxyfluorescein dye-labeled TaqMan probe, P-PDGF12, and a reverse primer, AS-PDGF12 (RQ primers/probe). C: One of the patient’s specific FIP1L1-PDGFRA sequences represented at the mRNA and protein levels. The fusion gene conserved an open reading frame.

    Journal: The Journal of molecular diagnostics : JMD

    Article Title: A Case of FIP1L1-PDGFRA-Positive Chronic Eosinophilic Leukemia with a Rare FIP1L1 Breakpoint

    doi: 10.2353/jmoldx.2007.060196

    Figure Lengend Snippet: Molecular analysis of the patient’s FIP1L1-PDGFRA fusion gene. A: Maps of FIP1L1 and PDGFRA genes. Boxes represent exons according to the LeukemiaNet exons numbering ( FIP1L1 , Ensembl Gene ID: ENSG00000145216; PDGFRA , ENSG00000134853). Numbers below the boxes indicate exon sizes in bp. The previously described FIP1L1 breakpoint region is indicated, encompassing introns 8 to 13, whereas PDGFRA breakpoint is invariably located in exon 12. The white arrow indicates our patient’s FIP1L1 intron 16-peculiar breakpoint. B: Patient’s specific FIP1L1-PDGFRA fusion gene juxtaposing a short portion of FIP1L1 intron 16 (33 bp) to PDGFRA exon 12. Primers and probes used in our assay are indicated. The diagnosis qualitative RT-PCR was performed with the primer pairs FIP1L1-F4 and PDGFRA-R1 for the first round and FIP1L1-F5 combined with PDGFRA-R2 for the second round (Diagnostic primers). Patient’s specific primers used for the qualitative follow-up were FIP1L1S15 and PDGFRAAS13 (Patient’s specific primers). For the quantitative RQ RT-PCR, exon 16 forward primer FIP1L1E16T was used in conjunction with a carboxyfluorescein dye-labeled TaqMan probe, P-PDGF12, and a reverse primer, AS-PDGF12 (RQ primers/probe). C: One of the patient’s specific FIP1L1-PDGFRA sequences represented at the mRNA and protein levels. The fusion gene conserved an open reading frame.

    Article Snippet: PCR products were detected using the TaqMan chemistry (Applied Biosystems).

    Techniques: Reverse Transcription Polymerase Chain Reaction, Diagnostic Assay, Labeling