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    Qiagen sensiscript rt
    Enrichment of high molecular weight cDNAs. LNCaP RNA was diluted and used in RT reactions as suggested by RNAimage Kit (GenHunter) but using three different reverse transcriptases and oligo dT primer HT11-C. Primers HT11-C and H-AP5 were used to generate cDNA pools and Taq polymerase was used in the PCR step. The resulting cDNAs at high ( A ) or low ( B ) molecular weight fragments generated after DD are shown. To determine the sensitivity of <t>Sensiscript,</t> additional LNCaP RNA was serially diluted and RT reactions were achieved using Sensiscript and MMLV reverse transcriptases. HT11-C primer and H-AP3 were used instead of the H-AP5 primer to generate cDNA pools. Taq polymerase was used in the amplification step. Under these conditions, the limit of detection of Sensiscript was 0.2 ng of total RNA in the RT reaction.
    Sensiscript Rt, supplied by Qiagen, used in various techniques. Bioz Stars score: 99/100, based on 517 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sensiscript rt/product/Qiagen
    Average 99 stars, based on 517 article reviews
    Price from $9.99 to $1999.99
    sensiscript rt - by Bioz Stars, 2020-07
    99/100 stars
      Buy from Supplier

    92
    Promega sensiscript rt kit
    Enrichment of high molecular weight cDNAs. LNCaP RNA was diluted and used in RT reactions as suggested by RNAimage Kit (GenHunter) but using three different reverse transcriptases and oligo dT primer HT11-C. Primers HT11-C and H-AP5 were used to generate cDNA pools and Taq polymerase was used in the PCR step. The resulting cDNAs at high ( A ) or low ( B ) molecular weight fragments generated after DD are shown. To determine the sensitivity of <t>Sensiscript,</t> additional LNCaP RNA was serially diluted and RT reactions were achieved using Sensiscript and MMLV reverse transcriptases. HT11-C primer and H-AP3 were used instead of the H-AP5 primer to generate cDNA pools. Taq polymerase was used in the amplification step. Under these conditions, the limit of detection of Sensiscript was 0.2 ng of total RNA in the RT reaction.
    Sensiscript Rt Kit, supplied by Promega, used in various techniques. Bioz Stars score: 92/100, based on 22 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/sensiscript rt kit/product/Promega
    Average 92 stars, based on 22 article reviews
    Price from $9.99 to $1999.99
    sensiscript rt kit - by Bioz Stars, 2020-07
    92/100 stars
      Buy from Supplier

    Image Search Results


    Enrichment of high molecular weight cDNAs. LNCaP RNA was diluted and used in RT reactions as suggested by RNAimage Kit (GenHunter) but using three different reverse transcriptases and oligo dT primer HT11-C. Primers HT11-C and H-AP5 were used to generate cDNA pools and Taq polymerase was used in the PCR step. The resulting cDNAs at high ( A ) or low ( B ) molecular weight fragments generated after DD are shown. To determine the sensitivity of Sensiscript, additional LNCaP RNA was serially diluted and RT reactions were achieved using Sensiscript and MMLV reverse transcriptases. HT11-C primer and H-AP3 were used instead of the H-AP5 primer to generate cDNA pools. Taq polymerase was used in the amplification step. Under these conditions, the limit of detection of Sensiscript was 0.2 ng of total RNA in the RT reaction.

    Journal: Nucleic Acids Research

    Article Title: Identification of differentially expressed genes from limited amounts of RNA

    doi:

    Figure Lengend Snippet: Enrichment of high molecular weight cDNAs. LNCaP RNA was diluted and used in RT reactions as suggested by RNAimage Kit (GenHunter) but using three different reverse transcriptases and oligo dT primer HT11-C. Primers HT11-C and H-AP5 were used to generate cDNA pools and Taq polymerase was used in the PCR step. The resulting cDNAs at high ( A ) or low ( B ) molecular weight fragments generated after DD are shown. To determine the sensitivity of Sensiscript, additional LNCaP RNA was serially diluted and RT reactions were achieved using Sensiscript and MMLV reverse transcriptases. HT11-C primer and H-AP3 were used instead of the H-AP5 primer to generate cDNA pools. Taq polymerase was used in the amplification step. Under these conditions, the limit of detection of Sensiscript was 0.2 ng of total RNA in the RT reaction.

    Article Snippet: Reactions containing 2.0 µl of Omniscript or Sensiscript RT (Qiagen, Santa Clarita, CA), 2.0 µl 10× RT buffer, 2.0 µl 5 mM dNTP (final concentration of 500 µM), 2.0 µl of 10 µM H-T11 C (final concentration of 1 µM), 1.0 µl RNase inhibitor (10 U/µl) and 10.0 µl DEPC-treated water were incubated at 37°C for 60 min and then at 93°C for 5 min.

    Techniques: Molecular Weight, Polymerase Chain Reaction, Generated, Amplification

    CRV substitutions inhibited mRNA capping. (A) Schematic diagram of the 3' end of the minigenome. The initiation sites within the le promoter (+1, +3) and at the first gs (+45) are indicated with green arrows, and the positive-sense small le (sm le), CAT 1 mRNA, and antigenome RNAs that are produced from this region of the minigenome are shown below in blue. The regions contained within the negative sense 56–75 and 91–113 primers and probes are indicated in red. (B) Primer extension analysis of RNA generated by wt and variant L proteins using the 56–75 primer and Sensiscript reverse transcriptase. Lane 1 is a marker prepared by end-labeling a 31 nt DNA oligonucleotide corresponding in sequence to the primer extension product representing initiation at the first gs signal (M 45–75). (C) Quantification of RNA initiated at the gs signal based on replicates of the experiments shown in panel B. The data are normalized to a mean of the two wt samples included in each experiment at 100% and the bars show the mean and standard deviation for three independent experiments. (D) Primer extension analysis of the RNA generated by wt and the R1339A and H1338A/R1339A variant polymerases using the 56–75 primer and Thermoscript reverse transcriptase. RNAs were treated with buffer only, or pyrophosphatase in buffer, as indicated by -/+ symbols. (E and F) Primer extension analysis of RNA generated by wt and variant L proteins, performed with Thermoscript reverse transcriptase with either the 56–75 (E), or 91–113 primer (F). (G) Northern blot analysis of small RNA transcripts produced from the minigenome. The RNA was migrated on a 6% urea-acrylamide gel and detected with an end-labeled 56–75 oligonucleotide probe.

    Journal: PLoS Pathogens

    Article Title: RNA elongation by respiratory syncytial virus polymerase is calibrated by conserved region V

    doi: 10.1371/journal.ppat.1006803

    Figure Lengend Snippet: CRV substitutions inhibited mRNA capping. (A) Schematic diagram of the 3' end of the minigenome. The initiation sites within the le promoter (+1, +3) and at the first gs (+45) are indicated with green arrows, and the positive-sense small le (sm le), CAT 1 mRNA, and antigenome RNAs that are produced from this region of the minigenome are shown below in blue. The regions contained within the negative sense 56–75 and 91–113 primers and probes are indicated in red. (B) Primer extension analysis of RNA generated by wt and variant L proteins using the 56–75 primer and Sensiscript reverse transcriptase. Lane 1 is a marker prepared by end-labeling a 31 nt DNA oligonucleotide corresponding in sequence to the primer extension product representing initiation at the first gs signal (M 45–75). (C) Quantification of RNA initiated at the gs signal based on replicates of the experiments shown in panel B. The data are normalized to a mean of the two wt samples included in each experiment at 100% and the bars show the mean and standard deviation for three independent experiments. (D) Primer extension analysis of the RNA generated by wt and the R1339A and H1338A/R1339A variant polymerases using the 56–75 primer and Thermoscript reverse transcriptase. RNAs were treated with buffer only, or pyrophosphatase in buffer, as indicated by -/+ symbols. (E and F) Primer extension analysis of RNA generated by wt and variant L proteins, performed with Thermoscript reverse transcriptase with either the 56–75 (E), or 91–113 primer (F). (G) Northern blot analysis of small RNA transcripts produced from the minigenome. The RNA was migrated on a 6% urea-acrylamide gel and detected with an end-labeled 56–75 oligonucleotide probe.

    Article Snippet: RNA samples were reverse transcribed at 37°C using the Sensiscript RT kit (Qiagen) and radiolabeled primers.

    Techniques: Produced, Generated, Variant Assay, Marker, End Labeling, Sequencing, Standard Deviation, Northern Blot, Acrylamide Gel Assay, Labeling

    BI-D inhibited transcription by preventing release of small le RNA and caused accumulation of abortive RNAs. (A) Schematic diagram of the 3' end of the transcription-competent minigenome. The RNA products are shown in blue and the regions contained within the primers and probes are indicated in red. (B and C) Primer extension analysis of RNAs produced from the minigenome in the presence of varying concentrations of BI-D, using Sensiscript reverse transcriptase and primers 56–75 (B) and 15–39 (C). (D, E) Primer extension analysis of RNAs with Thermoscript reverse transcriptase and either the 56–75 (D) or 91–113 primer (E). In panels B, D, and E, lane 1 shows markers of oligonucleotides representing the primer extension products from the respective initiation sites; in panel C, lanes 1 and 2 show markers. Note that in panels C-E, the markers were migrated on the same gels as the adjacent samples, but a different exposure was required to visualize them. (F, G) Quantification of replicates of the experiments shown in panels B and C. Panel F shows levels of RNA initiated at +1, +3, and gs detected by primer 56–75. Panel G shows levels of RNA initiated at +1 and +3, detected by primer extension with primer 15–39. The +1 and gs products in panel F and the data in panel G were normalized to the—BI-D control in each experiment, which was set to 100%. In panel F, because the +3 products analyzed with primer 56–75 were at the level of background in the absence of BI-D, the +3 products were normalized to the value obtained with 800 nM BI-D, which was set to 100%. The bars show the mean and standard deviation of three independent experiments. (H, I) Northern blot analysis of small RNAs produced from the transcription-competent minigenomes by L protein in the absence or presence of BI-D. (J) Comparison of the sizes of abortive RNAs produced by a capping-deficient L variant (R1339) or by wt L protein in the presence of 800 nM BI-D. In H-J, RNA was migrated on 6% urea-acrylamide gels and detected with the indicated probe. The ladders are described in materials and methods. The 26 nt marker was based on the migration of the bromophenol blue (BB) dye on a 6% gel.

    Journal: PLoS Pathogens

    Article Title: RNA elongation by respiratory syncytial virus polymerase is calibrated by conserved region V

    doi: 10.1371/journal.ppat.1006803

    Figure Lengend Snippet: BI-D inhibited transcription by preventing release of small le RNA and caused accumulation of abortive RNAs. (A) Schematic diagram of the 3' end of the transcription-competent minigenome. The RNA products are shown in blue and the regions contained within the primers and probes are indicated in red. (B and C) Primer extension analysis of RNAs produced from the minigenome in the presence of varying concentrations of BI-D, using Sensiscript reverse transcriptase and primers 56–75 (B) and 15–39 (C). (D, E) Primer extension analysis of RNAs with Thermoscript reverse transcriptase and either the 56–75 (D) or 91–113 primer (E). In panels B, D, and E, lane 1 shows markers of oligonucleotides representing the primer extension products from the respective initiation sites; in panel C, lanes 1 and 2 show markers. Note that in panels C-E, the markers were migrated on the same gels as the adjacent samples, but a different exposure was required to visualize them. (F, G) Quantification of replicates of the experiments shown in panels B and C. Panel F shows levels of RNA initiated at +1, +3, and gs detected by primer 56–75. Panel G shows levels of RNA initiated at +1 and +3, detected by primer extension with primer 15–39. The +1 and gs products in panel F and the data in panel G were normalized to the—BI-D control in each experiment, which was set to 100%. In panel F, because the +3 products analyzed with primer 56–75 were at the level of background in the absence of BI-D, the +3 products were normalized to the value obtained with 800 nM BI-D, which was set to 100%. The bars show the mean and standard deviation of three independent experiments. (H, I) Northern blot analysis of small RNAs produced from the transcription-competent minigenomes by L protein in the absence or presence of BI-D. (J) Comparison of the sizes of abortive RNAs produced by a capping-deficient L variant (R1339) or by wt L protein in the presence of 800 nM BI-D. In H-J, RNA was migrated on 6% urea-acrylamide gels and detected with the indicated probe. The ladders are described in materials and methods. The 26 nt marker was based on the migration of the bromophenol blue (BB) dye on a 6% gel.

    Article Snippet: RNA samples were reverse transcribed at 37°C using the Sensiscript RT kit (Qiagen) and radiolabeled primers.

    Techniques: Produced, Standard Deviation, Northern Blot, Variant Assay, Marker, Migration