stem-loop rt primer Search Results


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  • 99
    Thermo Fisher stem looped rt primers
    Stem Looped Rt Primers, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 61 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    97
    Millipore tsrna specific stem loop rt primers
    Tsrna Specific Stem Loop Rt Primers, supplied by Millipore, used in various techniques. Bioz Stars score: 97/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Kaneka Corp stem loop rt primer
    Stem Loop Rt Primer, supplied by Kaneka Corp, used in various techniques. Bioz Stars score: 92/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Thermo Fisher stem loop rt primer
    Validation of the <t>stem-loop</t> <t>RT-PCR</t> miRNA assays. (A) Genomic location of the JCPyV/BKPyV encoded miRNAs. (B) Graphical representation of a pre-miRNA as precursor of the mature 5p and 3p miRNAs generated upon cleavage by Dicer (C) Sequence comparison of the three miRNAs investigated. (D-F) The assay linearity and specificity was evaluated with dilution series of three synthetic miRNAs, miR-J1-5p, miR-J1a-5p and miR-B1-5p. Each dilution series was analyzed using the three miRNA assays. (G) The assay readings of miR-J1-5p by miR-J1-5p assay, miR-J1a-5p by miR-J1a-5p assay, and miR-B1-5p by miR-B1-5p assay at concentration levels of 2.10 4 copies/μL extract were used as the relative standards (100%) for the analysis of assay cross-reactivity.
    Stem Loop Rt Primer, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 1443 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/stem loop rt primer/product/Thermo Fisher
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    92
    Ribobio stem loop rt primer
    Validation of the <t>stem-loop</t> <t>RT-PCR</t> miRNA assays. (A) Genomic location of the JCPyV/BKPyV encoded miRNAs. (B) Graphical representation of a pre-miRNA as precursor of the mature 5p and 3p miRNAs generated upon cleavage by Dicer (C) Sequence comparison of the three miRNAs investigated. (D-F) The assay linearity and specificity was evaluated with dilution series of three synthetic miRNAs, miR-J1-5p, miR-J1a-5p and miR-B1-5p. Each dilution series was analyzed using the three miRNA assays. (G) The assay readings of miR-J1-5p by miR-J1-5p assay, miR-J1a-5p by miR-J1a-5p assay, and miR-B1-5p by miR-B1-5p assay at concentration levels of 2.10 4 copies/μL extract were used as the relative standards (100%) for the analysis of assay cross-reactivity.
    Stem Loop Rt Primer, supplied by Ribobio, used in various techniques. Bioz Stars score: 92/100, based on 51 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    GenScript stem loop rt primers
    Validation of the <t>stem-loop</t> <t>RT-PCR</t> miRNA assays. (A) Genomic location of the JCPyV/BKPyV encoded miRNAs. (B) Graphical representation of a pre-miRNA as precursor of the mature 5p and 3p miRNAs generated upon cleavage by Dicer (C) Sequence comparison of the three miRNAs investigated. (D-F) The assay linearity and specificity was evaluated with dilution series of three synthetic miRNAs, miR-J1-5p, miR-J1a-5p and miR-B1-5p. Each dilution series was analyzed using the three miRNA assays. (G) The assay readings of miR-J1-5p by miR-J1-5p assay, miR-J1a-5p by miR-J1a-5p assay, and miR-B1-5p by miR-B1-5p assay at concentration levels of 2.10 4 copies/μL extract were used as the relative standards (100%) for the analysis of assay cross-reactivity.
    Stem Loop Rt Primers, supplied by GenScript, used in various techniques. Bioz Stars score: 93/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    GenePharma Company stem loop rt primer
    Validation of the <t>stem-loop</t> <t>RT-PCR</t> miRNA assays. (A) Genomic location of the JCPyV/BKPyV encoded miRNAs. (B) Graphical representation of a pre-miRNA as precursor of the mature 5p and 3p miRNAs generated upon cleavage by Dicer (C) Sequence comparison of the three miRNAs investigated. (D-F) The assay linearity and specificity was evaluated with dilution series of three synthetic miRNAs, miR-J1-5p, miR-J1a-5p and miR-B1-5p. Each dilution series was analyzed using the three miRNA assays. (G) The assay readings of miR-J1-5p by miR-J1-5p assay, miR-J1a-5p by miR-J1a-5p assay, and miR-B1-5p by miR-B1-5p assay at concentration levels of 2.10 4 copies/μL extract were used as the relative standards (100%) for the analysis of assay cross-reactivity.
    Stem Loop Rt Primer, supplied by GenePharma Company, used in various techniques. Bioz Stars score: 90/100, based on 35 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    MiRXES stem loop rt primer pools
    Validation of the <t>stem-loop</t> <t>RT-PCR</t> miRNA assays. (A) Genomic location of the JCPyV/BKPyV encoded miRNAs. (B) Graphical representation of a pre-miRNA as precursor of the mature 5p and 3p miRNAs generated upon cleavage by Dicer (C) Sequence comparison of the three miRNAs investigated. (D-F) The assay linearity and specificity was evaluated with dilution series of three synthetic miRNAs, miR-J1-5p, miR-J1a-5p and miR-B1-5p. Each dilution series was analyzed using the three miRNA assays. (G) The assay readings of miR-J1-5p by miR-J1-5p assay, miR-J1a-5p by miR-J1a-5p assay, and miR-B1-5p by miR-B1-5p assay at concentration levels of 2.10 4 copies/μL extract were used as the relative standards (100%) for the analysis of assay cross-reactivity.
    Stem Loop Rt Primer Pools, supplied by MiRXES, used in various techniques. Bioz Stars score: 93/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Thermo Fisher megaplex stem loop rt primers
    Validation of the <t>stem-loop</t> <t>RT-PCR</t> miRNA assays. (A) Genomic location of the JCPyV/BKPyV encoded miRNAs. (B) Graphical representation of a pre-miRNA as precursor of the mature 5p and 3p miRNAs generated upon cleavage by Dicer (C) Sequence comparison of the three miRNAs investigated. (D-F) The assay linearity and specificity was evaluated with dilution series of three synthetic miRNAs, miR-J1-5p, miR-J1a-5p and miR-B1-5p. Each dilution series was analyzed using the three miRNA assays. (G) The assay readings of miR-J1-5p by miR-J1-5p assay, miR-J1a-5p by miR-J1a-5p assay, and miR-B1-5p by miR-B1-5p assay at concentration levels of 2.10 4 copies/μL extract were used as the relative standards (100%) for the analysis of assay cross-reactivity.
    Megaplex Stem Loop Rt Primers, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/megaplex stem loop rt primers/product/Thermo Fisher
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    88
    Ribobio stem loop rt pcr primers
    MiR-10b promotes autophagy by silencing DAZAP1 expression in ESCC. (A) Venn diagram of potential candidate target genes of miR-10b by integrating the results of the algorithms TargetScan, PICTAR, Micro-RNA and MiRDB. (B) <t>qRT-PCR</t> validation of the nine potential target genes of miR-10b in KYSE450 and KYSE510 cells transfected with either miR-10b mimics or NC RNA. (C) miR-10b could significantly inhibit DAZAP1 protein and mRNA expression in ESCC celllines. (D) Schematic constructions of pGL3-DAZAP1 and pGL3-Mut10b. (E) pGL3-DAZAP1 and pGL3-Mut10b were co-transfected into KYSE450 and KYSE510 cells with miR-10b mimics or NC RNA. Luciferase activity was detected at 48h after transfection and normalized relative to the Renilla luciferase expression. Inhibition effects of miR-10b mimics on pGL3-DAZAP1 or pGL3-Mut10b were showed. (F, G) Immunoblot results of extracts from non-starved or starved KYSE450 and KYSE510 cells. Silencing DAZAP1 expression (siDAZ1-1 and siDAZ1-2) increased starvation-induced conversion of LC3B-I to LC3B-II and accelerated rapamycin-induced SQSTM1 degradation in both ESCC celllines. Over-expressed DAZAP1 suppressed conversion of LC3B-I to LC3B-II and down-regulation of SQSTM1 in KYSE450 and KYSE510 cells. (H) DAZAP1 inhibited starvation-induced GFP-LC3 LC3B + autophagosomes formation in both ESCC celllines. The number of LC3 punctae in cells of each group was calculated from 3 random fields, and at least 30 cells were chosen. Autophagy was assessed under non-starved (STV-) or starved (STV+) conditions. The difference between two groups was calculated using Student's t test (assuming Gaussian distributions) or Wilcoxon Signed Rank Test (not assuming Gaussian distributions). All results of the mean of triplicate assays with standard deviation are presented. * P
    Stem Loop Rt Pcr Primers, supplied by Ribobio, used in various techniques. Bioz Stars score: 88/100, based on 27 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher stem loop commercial rt primers
    MiR-10b promotes autophagy by silencing DAZAP1 expression in ESCC. (A) Venn diagram of potential candidate target genes of miR-10b by integrating the results of the algorithms TargetScan, PICTAR, Micro-RNA and MiRDB. (B) <t>qRT-PCR</t> validation of the nine potential target genes of miR-10b in KYSE450 and KYSE510 cells transfected with either miR-10b mimics or NC RNA. (C) miR-10b could significantly inhibit DAZAP1 protein and mRNA expression in ESCC celllines. (D) Schematic constructions of pGL3-DAZAP1 and pGL3-Mut10b. (E) pGL3-DAZAP1 and pGL3-Mut10b were co-transfected into KYSE450 and KYSE510 cells with miR-10b mimics or NC RNA. Luciferase activity was detected at 48h after transfection and normalized relative to the Renilla luciferase expression. Inhibition effects of miR-10b mimics on pGL3-DAZAP1 or pGL3-Mut10b were showed. (F, G) Immunoblot results of extracts from non-starved or starved KYSE450 and KYSE510 cells. Silencing DAZAP1 expression (siDAZ1-1 and siDAZ1-2) increased starvation-induced conversion of LC3B-I to LC3B-II and accelerated rapamycin-induced SQSTM1 degradation in both ESCC celllines. Over-expressed DAZAP1 suppressed conversion of LC3B-I to LC3B-II and down-regulation of SQSTM1 in KYSE450 and KYSE510 cells. (H) DAZAP1 inhibited starvation-induced GFP-LC3 LC3B + autophagosomes formation in both ESCC celllines. The number of LC3 punctae in cells of each group was calculated from 3 random fields, and at least 30 cells were chosen. Autophagy was assessed under non-starved (STV-) or starved (STV+) conditions. The difference between two groups was calculated using Student's t test (assuming Gaussian distributions) or Wilcoxon Signed Rank Test (not assuming Gaussian distributions). All results of the mean of triplicate assays with standard deviation are presented. * P
    Stem Loop Commercial Rt Primers, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/stem loop commercial rt primers/product/Thermo Fisher
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    88
    Thermo Fisher stem loop shaped rt primer taqman assays
    MiR-10b promotes autophagy by silencing DAZAP1 expression in ESCC. (A) Venn diagram of potential candidate target genes of miR-10b by integrating the results of the algorithms TargetScan, PICTAR, Micro-RNA and MiRDB. (B) <t>qRT-PCR</t> validation of the nine potential target genes of miR-10b in KYSE450 and KYSE510 cells transfected with either miR-10b mimics or NC RNA. (C) miR-10b could significantly inhibit DAZAP1 protein and mRNA expression in ESCC celllines. (D) Schematic constructions of pGL3-DAZAP1 and pGL3-Mut10b. (E) pGL3-DAZAP1 and pGL3-Mut10b were co-transfected into KYSE450 and KYSE510 cells with miR-10b mimics or NC RNA. Luciferase activity was detected at 48h after transfection and normalized relative to the Renilla luciferase expression. Inhibition effects of miR-10b mimics on pGL3-DAZAP1 or pGL3-Mut10b were showed. (F, G) Immunoblot results of extracts from non-starved or starved KYSE450 and KYSE510 cells. Silencing DAZAP1 expression (siDAZ1-1 and siDAZ1-2) increased starvation-induced conversion of LC3B-I to LC3B-II and accelerated rapamycin-induced SQSTM1 degradation in both ESCC celllines. Over-expressed DAZAP1 suppressed conversion of LC3B-I to LC3B-II and down-regulation of SQSTM1 in KYSE450 and KYSE510 cells. (H) DAZAP1 inhibited starvation-induced GFP-LC3 LC3B + autophagosomes formation in both ESCC celllines. The number of LC3 punctae in cells of each group was calculated from 3 random fields, and at least 30 cells were chosen. Autophagy was assessed under non-starved (STV-) or starved (STV+) conditions. The difference between two groups was calculated using Student's t test (assuming Gaussian distributions) or Wilcoxon Signed Rank Test (not assuming Gaussian distributions). All results of the mean of triplicate assays with standard deviation are presented. * P
    Stem Loop Shaped Rt Primer Taqman Assays, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 88/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Sangon Biotech mirna specific stem loop rt primer
    MiR-10b promotes autophagy by silencing DAZAP1 expression in ESCC. (A) Venn diagram of potential candidate target genes of miR-10b by integrating the results of the algorithms TargetScan, PICTAR, Micro-RNA and MiRDB. (B) <t>qRT-PCR</t> validation of the nine potential target genes of miR-10b in KYSE450 and KYSE510 cells transfected with either miR-10b mimics or NC RNA. (C) miR-10b could significantly inhibit DAZAP1 protein and mRNA expression in ESCC celllines. (D) Schematic constructions of pGL3-DAZAP1 and pGL3-Mut10b. (E) pGL3-DAZAP1 and pGL3-Mut10b were co-transfected into KYSE450 and KYSE510 cells with miR-10b mimics or NC RNA. Luciferase activity was detected at 48h after transfection and normalized relative to the Renilla luciferase expression. Inhibition effects of miR-10b mimics on pGL3-DAZAP1 or pGL3-Mut10b were showed. (F, G) Immunoblot results of extracts from non-starved or starved KYSE450 and KYSE510 cells. Silencing DAZAP1 expression (siDAZ1-1 and siDAZ1-2) increased starvation-induced conversion of LC3B-I to LC3B-II and accelerated rapamycin-induced SQSTM1 degradation in both ESCC celllines. Over-expressed DAZAP1 suppressed conversion of LC3B-I to LC3B-II and down-regulation of SQSTM1 in KYSE450 and KYSE510 cells. (H) DAZAP1 inhibited starvation-induced GFP-LC3 LC3B + autophagosomes formation in both ESCC celllines. The number of LC3 punctae in cells of each group was calculated from 3 random fields, and at least 30 cells were chosen. Autophagy was assessed under non-starved (STV-) or starved (STV+) conditions. The difference between two groups was calculated using Student's t test (assuming Gaussian distributions) or Wilcoxon Signed Rank Test (not assuming Gaussian distributions). All results of the mean of triplicate assays with standard deviation are presented. * P
    Mirna Specific Stem Loop Rt Primer, supplied by Sangon Biotech, used in various techniques. Bioz Stars score: 92/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Thermo Fisher 5x mir 34a stem loop rt primer
    MiR-10b promotes autophagy by silencing DAZAP1 expression in ESCC. (A) Venn diagram of potential candidate target genes of miR-10b by integrating the results of the algorithms TargetScan, PICTAR, Micro-RNA and MiRDB. (B) <t>qRT-PCR</t> validation of the nine potential target genes of miR-10b in KYSE450 and KYSE510 cells transfected with either miR-10b mimics or NC RNA. (C) miR-10b could significantly inhibit DAZAP1 protein and mRNA expression in ESCC celllines. (D) Schematic constructions of pGL3-DAZAP1 and pGL3-Mut10b. (E) pGL3-DAZAP1 and pGL3-Mut10b were co-transfected into KYSE450 and KYSE510 cells with miR-10b mimics or NC RNA. Luciferase activity was detected at 48h after transfection and normalized relative to the Renilla luciferase expression. Inhibition effects of miR-10b mimics on pGL3-DAZAP1 or pGL3-Mut10b were showed. (F, G) Immunoblot results of extracts from non-starved or starved KYSE450 and KYSE510 cells. Silencing DAZAP1 expression (siDAZ1-1 and siDAZ1-2) increased starvation-induced conversion of LC3B-I to LC3B-II and accelerated rapamycin-induced SQSTM1 degradation in both ESCC celllines. Over-expressed DAZAP1 suppressed conversion of LC3B-I to LC3B-II and down-regulation of SQSTM1 in KYSE450 and KYSE510 cells. (H) DAZAP1 inhibited starvation-induced GFP-LC3 LC3B + autophagosomes formation in both ESCC celllines. The number of LC3 punctae in cells of each group was calculated from 3 random fields, and at least 30 cells were chosen. Autophagy was assessed under non-starved (STV-) or starved (STV+) conditions. The difference between two groups was calculated using Student's t test (assuming Gaussian distributions) or Wilcoxon Signed Rank Test (not assuming Gaussian distributions). All results of the mean of triplicate assays with standard deviation are presented. * P
    5x Mir 34a Stem Loop Rt Primer, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 88/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/5x mir 34a stem loop rt primer/product/Thermo Fisher
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    89
    Ribobio mirna specific stem loop rt primers
    MiR-10b promotes autophagy by silencing DAZAP1 expression in ESCC. (A) Venn diagram of potential candidate target genes of miR-10b by integrating the results of the algorithms TargetScan, PICTAR, Micro-RNA and MiRDB. (B) <t>qRT-PCR</t> validation of the nine potential target genes of miR-10b in KYSE450 and KYSE510 cells transfected with either miR-10b mimics or NC RNA. (C) miR-10b could significantly inhibit DAZAP1 protein and mRNA expression in ESCC celllines. (D) Schematic constructions of pGL3-DAZAP1 and pGL3-Mut10b. (E) pGL3-DAZAP1 and pGL3-Mut10b were co-transfected into KYSE450 and KYSE510 cells with miR-10b mimics or NC RNA. Luciferase activity was detected at 48h after transfection and normalized relative to the Renilla luciferase expression. Inhibition effects of miR-10b mimics on pGL3-DAZAP1 or pGL3-Mut10b were showed. (F, G) Immunoblot results of extracts from non-starved or starved KYSE450 and KYSE510 cells. Silencing DAZAP1 expression (siDAZ1-1 and siDAZ1-2) increased starvation-induced conversion of LC3B-I to LC3B-II and accelerated rapamycin-induced SQSTM1 degradation in both ESCC celllines. Over-expressed DAZAP1 suppressed conversion of LC3B-I to LC3B-II and down-regulation of SQSTM1 in KYSE450 and KYSE510 cells. (H) DAZAP1 inhibited starvation-induced GFP-LC3 LC3B + autophagosomes formation in both ESCC celllines. The number of LC3 punctae in cells of each group was calculated from 3 random fields, and at least 30 cells were chosen. Autophagy was assessed under non-starved (STV-) or starved (STV+) conditions. The difference between two groups was calculated using Student's t test (assuming Gaussian distributions) or Wilcoxon Signed Rank Test (not assuming Gaussian distributions). All results of the mean of triplicate assays with standard deviation are presented. * P
    Mirna Specific Stem Loop Rt Primers, supplied by Ribobio, used in various techniques. Bioz Stars score: 89/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Validation of the stem-loop RT-PCR miRNA assays. (A) Genomic location of the JCPyV/BKPyV encoded miRNAs. (B) Graphical representation of a pre-miRNA as precursor of the mature 5p and 3p miRNAs generated upon cleavage by Dicer (C) Sequence comparison of the three miRNAs investigated. (D-F) The assay linearity and specificity was evaluated with dilution series of three synthetic miRNAs, miR-J1-5p, miR-J1a-5p and miR-B1-5p. Each dilution series was analyzed using the three miRNA assays. (G) The assay readings of miR-J1-5p by miR-J1-5p assay, miR-J1a-5p by miR-J1a-5p assay, and miR-B1-5p by miR-B1-5p assay at concentration levels of 2.10 4 copies/μL extract were used as the relative standards (100%) for the analysis of assay cross-reactivity.

    Journal: Virology Journal

    Article Title: Viral miRNAs in plasma and urine divulge JC polyomavirus infection

    doi: 10.1186/1743-422X-11-158

    Figure Lengend Snippet: Validation of the stem-loop RT-PCR miRNA assays. (A) Genomic location of the JCPyV/BKPyV encoded miRNAs. (B) Graphical representation of a pre-miRNA as precursor of the mature 5p and 3p miRNAs generated upon cleavage by Dicer (C) Sequence comparison of the three miRNAs investigated. (D-F) The assay linearity and specificity was evaluated with dilution series of three synthetic miRNAs, miR-J1-5p, miR-J1a-5p and miR-B1-5p. Each dilution series was analyzed using the three miRNA assays. (G) The assay readings of miR-J1-5p by miR-J1-5p assay, miR-J1a-5p by miR-J1a-5p assay, and miR-B1-5p by miR-B1-5p assay at concentration levels of 2.10 4 copies/μL extract were used as the relative standards (100%) for the analysis of assay cross-reactivity.

    Article Snippet: Three μL of total RNA (representing 20% of total RNA extract) or synthetic miRNA solution was reverse transcribed using the pooled RT stem-loop primers (Life Technologies), enabling miRNA specific cDNA synthesis.

    Techniques: Reverse Transcription Polymerase Chain Reaction, Generated, Sequencing, Concentration Assay

    MiR-10b promotes autophagy by silencing DAZAP1 expression in ESCC. (A) Venn diagram of potential candidate target genes of miR-10b by integrating the results of the algorithms TargetScan, PICTAR, Micro-RNA and MiRDB. (B) qRT-PCR validation of the nine potential target genes of miR-10b in KYSE450 and KYSE510 cells transfected with either miR-10b mimics or NC RNA. (C) miR-10b could significantly inhibit DAZAP1 protein and mRNA expression in ESCC celllines. (D) Schematic constructions of pGL3-DAZAP1 and pGL3-Mut10b. (E) pGL3-DAZAP1 and pGL3-Mut10b were co-transfected into KYSE450 and KYSE510 cells with miR-10b mimics or NC RNA. Luciferase activity was detected at 48h after transfection and normalized relative to the Renilla luciferase expression. Inhibition effects of miR-10b mimics on pGL3-DAZAP1 or pGL3-Mut10b were showed. (F, G) Immunoblot results of extracts from non-starved or starved KYSE450 and KYSE510 cells. Silencing DAZAP1 expression (siDAZ1-1 and siDAZ1-2) increased starvation-induced conversion of LC3B-I to LC3B-II and accelerated rapamycin-induced SQSTM1 degradation in both ESCC celllines. Over-expressed DAZAP1 suppressed conversion of LC3B-I to LC3B-II and down-regulation of SQSTM1 in KYSE450 and KYSE510 cells. (H) DAZAP1 inhibited starvation-induced GFP-LC3 LC3B + autophagosomes formation in both ESCC celllines. The number of LC3 punctae in cells of each group was calculated from 3 random fields, and at least 30 cells were chosen. Autophagy was assessed under non-starved (STV-) or starved (STV+) conditions. The difference between two groups was calculated using Student's t test (assuming Gaussian distributions) or Wilcoxon Signed Rank Test (not assuming Gaussian distributions). All results of the mean of triplicate assays with standard deviation are presented. * P

    Journal: Theranostics

    Article Title: Starvation-induced suppression of DAZAP1 by miR-10b integrates splicing control into TSC2-regulated oncogenic autophagy in esophageal squamous cell carcinoma

    doi: 10.7150/thno.43046

    Figure Lengend Snippet: MiR-10b promotes autophagy by silencing DAZAP1 expression in ESCC. (A) Venn diagram of potential candidate target genes of miR-10b by integrating the results of the algorithms TargetScan, PICTAR, Micro-RNA and MiRDB. (B) qRT-PCR validation of the nine potential target genes of miR-10b in KYSE450 and KYSE510 cells transfected with either miR-10b mimics or NC RNA. (C) miR-10b could significantly inhibit DAZAP1 protein and mRNA expression in ESCC celllines. (D) Schematic constructions of pGL3-DAZAP1 and pGL3-Mut10b. (E) pGL3-DAZAP1 and pGL3-Mut10b were co-transfected into KYSE450 and KYSE510 cells with miR-10b mimics or NC RNA. Luciferase activity was detected at 48h after transfection and normalized relative to the Renilla luciferase expression. Inhibition effects of miR-10b mimics on pGL3-DAZAP1 or pGL3-Mut10b were showed. (F, G) Immunoblot results of extracts from non-starved or starved KYSE450 and KYSE510 cells. Silencing DAZAP1 expression (siDAZ1-1 and siDAZ1-2) increased starvation-induced conversion of LC3B-I to LC3B-II and accelerated rapamycin-induced SQSTM1 degradation in both ESCC celllines. Over-expressed DAZAP1 suppressed conversion of LC3B-I to LC3B-II and down-regulation of SQSTM1 in KYSE450 and KYSE510 cells. (H) DAZAP1 inhibited starvation-induced GFP-LC3 LC3B + autophagosomes formation in both ESCC celllines. The number of LC3 punctae in cells of each group was calculated from 3 random fields, and at least 30 cells were chosen. Autophagy was assessed under non-starved (STV-) or starved (STV+) conditions. The difference between two groups was calculated using Student's t test (assuming Gaussian distributions) or Wilcoxon Signed Rank Test (not assuming Gaussian distributions). All results of the mean of triplicate assays with standard deviation are presented. * P

    Article Snippet: Human miRNAs and U6 were detected with their specific stem-loop RT-PCR primers (Ribobio, Guangzhou, China) , .

    Techniques: Expressing, Quantitative RT-PCR, Transfection, Luciferase, Activity Assay, Inhibition, Standard Deviation