total cellular rna Search Results


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  • 99
    Thermo Fisher total cellular rna isolation
    Total Cellular Rna Isolation, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 27 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore total cellular rna
    Analysis of intracellular and HIV-1 virion-associated and tRNA Glu . (A) Analysis of tRNA from <t>SupT1</t> cells. Total <t>RNA</t> from SupT1 cells was isolated as described in Materials and Methods. Following electrophoresis and transfer onto nitrocellulose membrane, equal amounts were probed with oligonucleotides specific for or tRNA Glu . Specificity controls included equal amounts (1 ng) of in vitro RNA transcription products; note that the in vitro transcript (xscript) for tRNA Glu contains extra nucleotides that retard the migration in the agarose gels used for this analysis. The densities of the probes reacting with the cognate tRNA for both and tRNA Glu were similar (i.e., the density of the probe with 1 ng of in vitro-transcribed versus the density of the tRNA Glu probe with 1 ng of in vitro-transcribed tRNA Glu ). The use of scanning densitometry to determine the cellular amounts of tRNA Glu and revealed an approximate 10:1 ratio. This analysis was repeated three times, and a representative blot is shown. The closed arrow denotes migration of full-length tRNA Lys or tRNA Glu . The open arrow denotes migration of the tRNA Glu in vitro transcript containing additional nucleotides from cloning of the tRNA gene into the transcription plasmid. (B) Analysis of tRNA from HIV-1 virions. Total virion RNA collected from viruses isolated from supernatant of SupT1 cells infected with HXB2 or HXB2(Glu Loop 2). HIV-1 virions were isolated by ultracentrifugation, and RNA was extracted and prepared as described in Materials and Methods. The blot was probed for tRNA Glu and as indicated. Lane 1, 1 μg of RNA isolated from virions; lane 2, 0.1 μg of RNA isolated from virions.
    Total Cellular Rna, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 1001 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Becton Dickinson total cellular rna
    Analysis of intracellular and HIV-1 virion-associated and tRNA Glu . (A) Analysis of tRNA from <t>SupT1</t> cells. Total <t>RNA</t> from SupT1 cells was isolated as described in Materials and Methods. Following electrophoresis and transfer onto nitrocellulose membrane, equal amounts were probed with oligonucleotides specific for or tRNA Glu . Specificity controls included equal amounts (1 ng) of in vitro RNA transcription products; note that the in vitro transcript (xscript) for tRNA Glu contains extra nucleotides that retard the migration in the agarose gels used for this analysis. The densities of the probes reacting with the cognate tRNA for both and tRNA Glu were similar (i.e., the density of the probe with 1 ng of in vitro-transcribed versus the density of the tRNA Glu probe with 1 ng of in vitro-transcribed tRNA Glu ). The use of scanning densitometry to determine the cellular amounts of tRNA Glu and revealed an approximate 10:1 ratio. This analysis was repeated three times, and a representative blot is shown. The closed arrow denotes migration of full-length tRNA Lys or tRNA Glu . The open arrow denotes migration of the tRNA Glu in vitro transcript containing additional nucleotides from cloning of the tRNA gene into the transcription plasmid. (B) Analysis of tRNA from HIV-1 virions. Total virion RNA collected from viruses isolated from supernatant of SupT1 cells infected with HXB2 or HXB2(Glu Loop 2). HIV-1 virions were isolated by ultracentrifugation, and RNA was extracted and prepared as described in Materials and Methods. The blot was probed for tRNA Glu and as indicated. Lane 1, 1 μg of RNA isolated from virions; lane 2, 0.1 μg of RNA isolated from virions.
    Total Cellular Rna, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 92/100, based on 39 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Thermo Fisher total cellular rna
    <t>cDNA</t> slot blots of <t>ER-RNA.</t> ( A ) Dilution series of cDNA obtained when r(CUG) 100 template was singly primed from upstream site versus multiply primed within the repeat tract by (CAG) 6+1 (RNA/DNA hybrid primers). ( B ) Quantification of cDNA slot blot in (A), (n = 3 for each condition). ( C ) cDNA slot blot analysis of total cellular RNA (2 μg) from DM1 versus non-DM1 cardiac tissue using all-DNA versus RNA/DNA hybrid primers. Both reactions in (C) used 3 dNTPs plus ddUTP. ( D ) cDNA slot blot analysis of total cellular RNA (2 μg) from healthy control or DM1 tibialis anterior muscle biopsy samples using all 4 dNTPs versus 3 dNTPs (dCTP, dATP, dGTP plus ddUTP chain terminator) in the RT reaction. The reactions in (D) used RNA/DNA hybrid primers.
    Total Cellular Rna, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 15859 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    MACHEREY NAGEL total cellular rna macherey nagel
    piggyBac transposase bioavailability. ( A ) Half-life of the PB <t>mRNA</t> . Cells were transfected with 187.5 ng of PB mRNA. Total <t>RNA</t> was extracted at indicated times, reverse transcribed and subjected to qPCR. 18S RNA served as an internal standard to normalize the data. ( B ) Persistence of PB pDNA after transfection . Cells were transfected with 187.5 ng of PB pDNA and plasmid rescue was performed at 0 to 20 days. Ampicillin-resistant colonies were selected to evaluate the persistence of the plasmid. ( C ) Half-life of the PB transposase (V5PB Tp) . Cells were transfected with 187.5 ng of PB mRNA, incubated 18 h to reach the peak of transposase expression and treated with cycloheximide (t0=100). Total protein extraction was done at the indicated times from t0. The transposase half-life was determined by Western-Blot and quantification was normalized to the endogenous actin protein.
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    TaKaRa total cellular rna
    Effects of I3C on the expression of cell-cycle and apoptosis regulatory proteins . Western blot analysis of HUT-102 and TL-OmI cells treated with I3C. (A) Cells were treated with I3C (100 μM) for the indicated periods. (B) Cells were treated with I3C at the indicated concentrations for 48 h. Total cellular proteins (20 μg per lane) were separated on SDS-polyacrylamide gels and transferred to the membrane. Protein levels were detected by Western blotting with antibodies directed against each protein. (C) Total <t>RNA</t> was extracted from HUT-102 and TL-OmI cells following treatment with I3C (100 μM) for 12 or 24 h. The mRNA expression of Tax and HBZ was analyzed by <t>RT-PCR</t> analysis. β-actin served as an internal control in the RT-PCR procedure. Representative data of three experiments with similar results.
    Total Cellular Rna, supplied by TaKaRa, used in various techniques. Bioz Stars score: 93/100, based on 772 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Tel Test Inc total cellular rna
    Homologous rat sequences can partially substitute for the intronic enhancer but not the silencer in intron 10 of human gene. Schematic diagram of human IR minigene chimeras with rat sequence substitutions is shown on left. White boxes represent human exons 10 and 11 separated by intron 10. Dotted lines indicate deletions in the intervening intron. Gray boxes indicate the homologous sequence from the rat Insr gene. Numbers below the introns indicate sizes of deletions. Numbers above the intron and gray boxes indicate length of human or rat sequence. The positions of the human intronic splicing enhancer ( ISE ) or silencer ( ISS ) are indicated in hIRB. These minigenes were transfected into HepG2 cells. Total <t>RNA</t> was isolated 48 h <t>post-transfection</t> and was subjected to RT-PCR analysis using primers specific to the transfected IR minigene mRNA. The mean ± S.E. for percent exon 11 inclusion is shown as a bar graph on the right . Results are derived from at least three independent experiments. Statistical significance was analyzed by ANOVA. Bars with the same letter are not significantly different ( p
    Total Cellular Rna, supplied by Tel Test Inc, used in various techniques. Bioz Stars score: 92/100, based on 236 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Arraystar total cellular rna
    Homologous rat sequences can partially substitute for the intronic enhancer but not the silencer in intron 10 of human gene. Schematic diagram of human IR minigene chimeras with rat sequence substitutions is shown on left. White boxes represent human exons 10 and 11 separated by intron 10. Dotted lines indicate deletions in the intervening intron. Gray boxes indicate the homologous sequence from the rat Insr gene. Numbers below the introns indicate sizes of deletions. Numbers above the intron and gray boxes indicate length of human or rat sequence. The positions of the human intronic splicing enhancer ( ISE ) or silencer ( ISS ) are indicated in hIRB. These minigenes were transfected into HepG2 cells. Total <t>RNA</t> was isolated 48 h <t>post-transfection</t> and was subjected to RT-PCR analysis using primers specific to the transfected IR minigene mRNA. The mean ± S.E. for percent exon 11 inclusion is shown as a bar graph on the right . Results are derived from at least three independent experiments. Statistical significance was analyzed by ANOVA. Bars with the same letter are not significantly different ( p
    Total Cellular Rna, supplied by Arraystar, used in various techniques. Bioz Stars score: 93/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Molecular Research Center inc total cellular rna
    Detection of <t>SHFV</t> <t>RNA</t> by RT-PCR in baboon sera. RNA isolated from 100 μl of baboon serum is analyzed by one-step RT-PCR. Pairs of primers are designed to amplify (A) nucleocapsid or (B) helicase regions of the SHFV genome. RNA extracted from an
    Total Cellular Rna, supplied by Molecular Research Center inc, used in various techniques. Bioz Stars score: 92/100, based on 707 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    iNtRON Biotechnology total cellular rna
    Detection of <t>SHFV</t> <t>RNA</t> by RT-PCR in baboon sera. RNA isolated from 100 μl of baboon serum is analyzed by one-step RT-PCR. Pairs of primers are designed to amplify (A) nucleocapsid or (B) helicase regions of the SHFV genome. RNA extracted from an
    Total Cellular Rna, supplied by iNtRON Biotechnology, used in various techniques. Bioz Stars score: 91/100, based on 79 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    AMS Biotechnology total cellular rna
    The Inflammatory Response Depends on the RNase Activity of <t>IRE1α</t> and on RIG-I, a Cytosolic Sensor of <t>RNA</t> Fragments
    Total Cellular Rna, supplied by AMS Biotechnology, used in various techniques. Bioz Stars score: 92/100, based on 20 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher totally rna total cellular rna isolation kit
    The Inflammatory Response Depends on the RNase Activity of <t>IRE1α</t> and on RIG-I, a Cytosolic Sensor of <t>RNA</t> Fragments
    Totally Rna Total Cellular Rna Isolation Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    FUJIFILM total cellular rna
    The Inflammatory Response Depends on the RNase Activity of <t>IRE1α</t> and on RIG-I, a Cytosolic Sensor of <t>RNA</t> Fragments
    Total Cellular Rna, supplied by FUJIFILM, used in various techniques. Bioz Stars score: 92/100, based on 58 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher additional human total cellular rna
    The Inflammatory Response Depends on the RNase Activity of <t>IRE1α</t> and on RIG-I, a Cytosolic Sensor of <t>RNA</t> Fragments
    Additional Human Total Cellular Rna, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TaKaRa human total cellular rna
    The Inflammatory Response Depends on the RNase Activity of <t>IRE1α</t> and on RIG-I, a Cytosolic Sensor of <t>RNA</t> Fragments
    Human Total Cellular Rna, supplied by TaKaRa, used in various techniques. Bioz Stars score: 99/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    TaKaRa cellular rna total cellular rna
    The Inflammatory Response Depends on the RNase Activity of <t>IRE1α</t> and on RIG-I, a Cytosolic Sensor of <t>RNA</t> Fragments
    Cellular Rna Total Cellular Rna, supplied by TaKaRa, used in various techniques. Bioz Stars score: 86/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Illumina Inc hiseq total cellular rna
    The Inflammatory Response Depends on the RNase Activity of <t>IRE1α</t> and on RIG-I, a Cytosolic Sensor of <t>RNA</t> Fragments
    Hiseq Total Cellular Rna, supplied by Illumina Inc, used in various techniques. Bioz Stars score: 92/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Omega Bio-tek total cellular rna isolation
    The Inflammatory Response Depends on the RNase Activity of <t>IRE1α</t> and on RIG-I, a Cytosolic Sensor of <t>RNA</t> Fragments
    Total Cellular Rna Isolation, supplied by Omega Bio-tek, used in various techniques. Bioz Stars score: 88/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher g total cellular rna
    The Inflammatory Response Depends on the RNase Activity of <t>IRE1α</t> and on RIG-I, a Cytosolic Sensor of <t>RNA</t> Fragments
    G Total Cellular Rna, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Stratagene qpcr mouse reference total cellular rna
    The Inflammatory Response Depends on the RNase Activity of <t>IRE1α</t> and on RIG-I, a Cytosolic Sensor of <t>RNA</t> Fragments
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    Image Search Results


    Analysis of intracellular and HIV-1 virion-associated and tRNA Glu . (A) Analysis of tRNA from SupT1 cells. Total RNA from SupT1 cells was isolated as described in Materials and Methods. Following electrophoresis and transfer onto nitrocellulose membrane, equal amounts were probed with oligonucleotides specific for or tRNA Glu . Specificity controls included equal amounts (1 ng) of in vitro RNA transcription products; note that the in vitro transcript (xscript) for tRNA Glu contains extra nucleotides that retard the migration in the agarose gels used for this analysis. The densities of the probes reacting with the cognate tRNA for both and tRNA Glu were similar (i.e., the density of the probe with 1 ng of in vitro-transcribed versus the density of the tRNA Glu probe with 1 ng of in vitro-transcribed tRNA Glu ). The use of scanning densitometry to determine the cellular amounts of tRNA Glu and revealed an approximate 10:1 ratio. This analysis was repeated three times, and a representative blot is shown. The closed arrow denotes migration of full-length tRNA Lys or tRNA Glu . The open arrow denotes migration of the tRNA Glu in vitro transcript containing additional nucleotides from cloning of the tRNA gene into the transcription plasmid. (B) Analysis of tRNA from HIV-1 virions. Total virion RNA collected from viruses isolated from supernatant of SupT1 cells infected with HXB2 or HXB2(Glu Loop 2). HIV-1 virions were isolated by ultracentrifugation, and RNA was extracted and prepared as described in Materials and Methods. The blot was probed for tRNA Glu and as indicated. Lane 1, 1 μg of RNA isolated from virions; lane 2, 0.1 μg of RNA isolated from virions.

    Journal: Journal of Virology

    Article Title: Probing the Importance of tRNA Anticodon: Human Immunodeficiency Virus Type 1 (HIV-1) RNA Genome Complementarity with an HIV-1 That Selects tRNAGlu for Replication

    doi: 10.1128/JVI.77.16.8756-8764.2003

    Figure Lengend Snippet: Analysis of intracellular and HIV-1 virion-associated and tRNA Glu . (A) Analysis of tRNA from SupT1 cells. Total RNA from SupT1 cells was isolated as described in Materials and Methods. Following electrophoresis and transfer onto nitrocellulose membrane, equal amounts were probed with oligonucleotides specific for or tRNA Glu . Specificity controls included equal amounts (1 ng) of in vitro RNA transcription products; note that the in vitro transcript (xscript) for tRNA Glu contains extra nucleotides that retard the migration in the agarose gels used for this analysis. The densities of the probes reacting with the cognate tRNA for both and tRNA Glu were similar (i.e., the density of the probe with 1 ng of in vitro-transcribed versus the density of the tRNA Glu probe with 1 ng of in vitro-transcribed tRNA Glu ). The use of scanning densitometry to determine the cellular amounts of tRNA Glu and revealed an approximate 10:1 ratio. This analysis was repeated three times, and a representative blot is shown. The closed arrow denotes migration of full-length tRNA Lys or tRNA Glu . The open arrow denotes migration of the tRNA Glu in vitro transcript containing additional nucleotides from cloning of the tRNA gene into the transcription plasmid. (B) Analysis of tRNA from HIV-1 virions. Total virion RNA collected from viruses isolated from supernatant of SupT1 cells infected with HXB2 or HXB2(Glu Loop 2). HIV-1 virions were isolated by ultracentrifugation, and RNA was extracted and prepared as described in Materials and Methods. The blot was probed for tRNA Glu and as indicated. Lane 1, 1 μg of RNA isolated from virions; lane 2, 0.1 μg of RNA isolated from virions.

    Article Snippet: Total cellular RNA was isolated from uninfected SupT1 cells by using Tri-reagent (Sigma) according to the manufacturer's instructions.

    Techniques: Isolation, Electrophoresis, In Vitro, Migration, Clone Assay, Plasmid Preparation, Infection

    cDNA slot blots of ER-RNA. ( A ) Dilution series of cDNA obtained when r(CUG) 100 template was singly primed from upstream site versus multiply primed within the repeat tract by (CAG) 6+1 (RNA/DNA hybrid primers). ( B ) Quantification of cDNA slot blot in (A), (n = 3 for each condition). ( C ) cDNA slot blot analysis of total cellular RNA (2 μg) from DM1 versus non-DM1 cardiac tissue using all-DNA versus RNA/DNA hybrid primers. Both reactions in (C) used 3 dNTPs plus ddUTP. ( D ) cDNA slot blot analysis of total cellular RNA (2 μg) from healthy control or DM1 tibialis anterior muscle biopsy samples using all 4 dNTPs versus 3 dNTPs (dCTP, dATP, dGTP plus ddUTP chain terminator) in the RT reaction. The reactions in (D) used RNA/DNA hybrid primers.

    Journal: Nucleic Acids Research

    Article Title: Detection of expanded RNA repeats using thermostable group II intron reverse transcriptase

    doi: 10.1093/nar/gkx867

    Figure Lengend Snippet: cDNA slot blots of ER-RNA. ( A ) Dilution series of cDNA obtained when r(CUG) 100 template was singly primed from upstream site versus multiply primed within the repeat tract by (CAG) 6+1 (RNA/DNA hybrid primers). ( B ) Quantification of cDNA slot blot in (A), (n = 3 for each condition). ( C ) cDNA slot blot analysis of total cellular RNA (2 μg) from DM1 versus non-DM1 cardiac tissue using all-DNA versus RNA/DNA hybrid primers. Both reactions in (C) used 3 dNTPs plus ddUTP. ( D ) cDNA slot blot analysis of total cellular RNA (2 μg) from healthy control or DM1 tibialis anterior muscle biopsy samples using all 4 dNTPs versus 3 dNTPs (dCTP, dATP, dGTP plus ddUTP chain terminator) in the RT reaction. The reactions in (D) used RNA/DNA hybrid primers.

    Article Snippet: Although our initial testing was performed using individual ER-RNAs in isolation, we used conditions designed to minimize off-target cDNA synthesis when later applied to total cellular RNA.

    Techniques: Dot Blot

    ER-RNA expression in transgenic mouse models of DM1. ( A ) cDNA slot blot of total cellular RNA from HSA LR quadriceps (Quads, 2 μg, three different mice) using Superscript-III (SS-III) or TGIRT-III. ( B ) cDNA slot blot of 2 μg total cellular RNA from quadriceps or tibialis anterior (TA) muscles from HSA LR and HSA XLR mice. Bottom-most wells in each column are from HSA SR or HSA NR mice, which do not express ER-RNA. ( C ) Relative amount of ER-RNA by cDNA slot blot in HSA LR and HSA XLR quadriceps and TA muscle (black bars, mean signal in HSA LR quadriceps set to 1), as compared to inferred ER-RNA level based on qRT-PCR (white bars, calculated by multiplying expression level x repeat length, with the mean product in HSA LR quadriceps set to 1). Results are based on n = 3 for HSA LR and HSA XLR mice. HSA transgene expression by qRT-PCR was normalized to general transcription factor 2b (Gtf2b) .

    Journal: Nucleic Acids Research

    Article Title: Detection of expanded RNA repeats using thermostable group II intron reverse transcriptase

    doi: 10.1093/nar/gkx867

    Figure Lengend Snippet: ER-RNA expression in transgenic mouse models of DM1. ( A ) cDNA slot blot of total cellular RNA from HSA LR quadriceps (Quads, 2 μg, three different mice) using Superscript-III (SS-III) or TGIRT-III. ( B ) cDNA slot blot of 2 μg total cellular RNA from quadriceps or tibialis anterior (TA) muscles from HSA LR and HSA XLR mice. Bottom-most wells in each column are from HSA SR or HSA NR mice, which do not express ER-RNA. ( C ) Relative amount of ER-RNA by cDNA slot blot in HSA LR and HSA XLR quadriceps and TA muscle (black bars, mean signal in HSA LR quadriceps set to 1), as compared to inferred ER-RNA level based on qRT-PCR (white bars, calculated by multiplying expression level x repeat length, with the mean product in HSA LR quadriceps set to 1). Results are based on n = 3 for HSA LR and HSA XLR mice. HSA transgene expression by qRT-PCR was normalized to general transcription factor 2b (Gtf2b) .

    Article Snippet: Although our initial testing was performed using individual ER-RNAs in isolation, we used conditions designed to minimize off-target cDNA synthesis when later applied to total cellular RNA.

    Techniques: RNA Expression, Transgenic Assay, Dot Blot, Mouse Assay, Quantitative RT-PCR, Expressing

    Reverse transcription across ER-RNA tracts using TGIRT-III or SS-III. ( A ) Diagram of ER-RNA showing HEX-labeled primer annealed 3′ of repeat tract and unlabeled primers tiled across the expanded repeat. ( B ) Representative chromatogram of cDNA generated by TGIRT-III from r(CUG) 100 in the presence or absence of intervening repeat primers. ( C ) Representative chromatogram of cDNA generated by TGIRT-III from r(GGGGCC) 40 , as in (B). ( D and E ) Representative chromatograms from r(CUG) 100 and r(GGGGCC) 40 templates using Superscript-III, as in (B) and (C).

    Journal: Nucleic Acids Research

    Article Title: Detection of expanded RNA repeats using thermostable group II intron reverse transcriptase

    doi: 10.1093/nar/gkx867

    Figure Lengend Snippet: Reverse transcription across ER-RNA tracts using TGIRT-III or SS-III. ( A ) Diagram of ER-RNA showing HEX-labeled primer annealed 3′ of repeat tract and unlabeled primers tiled across the expanded repeat. ( B ) Representative chromatogram of cDNA generated by TGIRT-III from r(CUG) 100 in the presence or absence of intervening repeat primers. ( C ) Representative chromatogram of cDNA generated by TGIRT-III from r(GGGGCC) 40 , as in (B). ( D and E ) Representative chromatograms from r(CUG) 100 and r(GGGGCC) 40 templates using Superscript-III, as in (B) and (C).

    Article Snippet: Although our initial testing was performed using individual ER-RNAs in isolation, we used conditions designed to minimize off-target cDNA synthesis when later applied to total cellular RNA.

    Techniques: Labeling, Generated

    Reverse transcription of enzymatically synthesized ER-RNAs. ( A ) Diagram of ER-RNA template, multiply primed with 5′ HEX end-labeled oligonucleotides. Representative chromatograms of HEX-labeled cDNA generated by TGIRT-III ( B–E ) or Superscript-III ( F–I ). Each trace represents analysis of a single cDNA synthesis.

    Journal: Nucleic Acids Research

    Article Title: Detection of expanded RNA repeats using thermostable group II intron reverse transcriptase

    doi: 10.1093/nar/gkx867

    Figure Lengend Snippet: Reverse transcription of enzymatically synthesized ER-RNAs. ( A ) Diagram of ER-RNA template, multiply primed with 5′ HEX end-labeled oligonucleotides. Representative chromatograms of HEX-labeled cDNA generated by TGIRT-III ( B–E ) or Superscript-III ( F–I ). Each trace represents analysis of a single cDNA synthesis.

    Article Snippet: Although our initial testing was performed using individual ER-RNAs in isolation, we used conditions designed to minimize off-target cDNA synthesis when later applied to total cellular RNA.

    Techniques: Synthesized, Labeling, Generated

    cDNA slot blot of DM1 and DM2 muscle biopsy samples and (GGGGCC) 160 -expressing cells. ( A ) cDNA slot blot of total cellular RNA (2 μg) from tibialis anterior (TA) biopsy samples of DM1 patients ( n = 3) or healthy controls, as compared to quadriceps muscle from HSA LR transgenic mice. The bottom-most wells in each column are HSA SR or healthy controls. ( B ) Relative quantification of (A), with the mean value from HSA LR mice set to 1. ( C ) cDNA slot blot of 2 μg total cellular RNA from TA biopsy samples from DM2 patients ( n = 6) or healthy controls ( n = 3, upper-most wells). Reverse transcription was primed with (CAGG) 4+2 RNA/DNA hybrid primer, as shown in table 1 . ( D ) Representative ER-cDNA slot blot of 2 μg total RNA from pLC19 (no repeat) versus pLC21 [expressing (GGGGCC) 160 ] cell lysates. Blots were run simultaneously, with an intervening slot removed for presentation.

    Journal: Nucleic Acids Research

    Article Title: Detection of expanded RNA repeats using thermostable group II intron reverse transcriptase

    doi: 10.1093/nar/gkx867

    Figure Lengend Snippet: cDNA slot blot of DM1 and DM2 muscle biopsy samples and (GGGGCC) 160 -expressing cells. ( A ) cDNA slot blot of total cellular RNA (2 μg) from tibialis anterior (TA) biopsy samples of DM1 patients ( n = 3) or healthy controls, as compared to quadriceps muscle from HSA LR transgenic mice. The bottom-most wells in each column are HSA SR or healthy controls. ( B ) Relative quantification of (A), with the mean value from HSA LR mice set to 1. ( C ) cDNA slot blot of 2 μg total cellular RNA from TA biopsy samples from DM2 patients ( n = 6) or healthy controls ( n = 3, upper-most wells). Reverse transcription was primed with (CAGG) 4+2 RNA/DNA hybrid primer, as shown in table 1 . ( D ) Representative ER-cDNA slot blot of 2 μg total RNA from pLC19 (no repeat) versus pLC21 [expressing (GGGGCC) 160 ] cell lysates. Blots were run simultaneously, with an intervening slot removed for presentation.

    Article Snippet: Although our initial testing was performed using individual ER-RNAs in isolation, we used conditions designed to minimize off-target cDNA synthesis when later applied to total cellular RNA.

    Techniques: Dot Blot, Expressing, Transgenic Assay, Mouse Assay

    Responsivity of cDNA slot blot and alternative splicing to treatment with antisense oligonucleotides (ASOs) in transgenic mice. ( A ) cDNA slot blot of 2 μg total RNA from quadriceps muscle (HSA LR and HSA XLR mice) or left ventricle (LC15 mice) treated with subcutaneous injections of saline ( n = 4) or ASO ( n = 4 in each group). Negative control samples from HSA SR quadriceps, HSA NR quadriceps, or wild-type (WT) heart are indicated. ( B ) Relative ER-RNA quantification by cDNA slot blot (black bars) versus qRT-PCR (white bars). Saline-treated groups set to 1 for both assays. ( C ) RT-PCR analysis of alternative splicing in saline- versus ASO-treated HSA LR quadriceps ( Serca1 and Clcn1 ) and LC15 heart tissue ( Tmem63b and Cacna1s ), as compared to wild-type controls. The mean ± S.D. values of ‘percent spliced in’ for saline versus ASO versus wild-type groups were Serca1 exon 22: 15 ± 2%, 94 ± 2%, 100 ± 0%; Clcn1 exon 7A: 31 ± 2%, 5 ± 1%, 4 ± 2%); Tmem63b exon 5: 46 ± 4%, 66 ± 2%, 84 ± 0%; Cacna1s exon 29: 38 ± 1%, 78 ± 1%, 95 ± 1%). ( D ) Quantification of (C). ANOVA of Serca1 exon 22 ( P = 2.2 × 10 −9 ), Clcn1 exon 7A ( P = 4.4 × 10 −6 ), Tmem63b exon 5 ( P = 3.1 × 10 −9 ), and Cacna1s ( P = 3.7 × 10 −9 ). *Student's t -test versus WT P

    Journal: Nucleic Acids Research

    Article Title: Detection of expanded RNA repeats using thermostable group II intron reverse transcriptase

    doi: 10.1093/nar/gkx867

    Figure Lengend Snippet: Responsivity of cDNA slot blot and alternative splicing to treatment with antisense oligonucleotides (ASOs) in transgenic mice. ( A ) cDNA slot blot of 2 μg total RNA from quadriceps muscle (HSA LR and HSA XLR mice) or left ventricle (LC15 mice) treated with subcutaneous injections of saline ( n = 4) or ASO ( n = 4 in each group). Negative control samples from HSA SR quadriceps, HSA NR quadriceps, or wild-type (WT) heart are indicated. ( B ) Relative ER-RNA quantification by cDNA slot blot (black bars) versus qRT-PCR (white bars). Saline-treated groups set to 1 for both assays. ( C ) RT-PCR analysis of alternative splicing in saline- versus ASO-treated HSA LR quadriceps ( Serca1 and Clcn1 ) and LC15 heart tissue ( Tmem63b and Cacna1s ), as compared to wild-type controls. The mean ± S.D. values of ‘percent spliced in’ for saline versus ASO versus wild-type groups were Serca1 exon 22: 15 ± 2%, 94 ± 2%, 100 ± 0%; Clcn1 exon 7A: 31 ± 2%, 5 ± 1%, 4 ± 2%); Tmem63b exon 5: 46 ± 4%, 66 ± 2%, 84 ± 0%; Cacna1s exon 29: 38 ± 1%, 78 ± 1%, 95 ± 1%). ( D ) Quantification of (C). ANOVA of Serca1 exon 22 ( P = 2.2 × 10 −9 ), Clcn1 exon 7A ( P = 4.4 × 10 −6 ), Tmem63b exon 5 ( P = 3.1 × 10 −9 ), and Cacna1s ( P = 3.7 × 10 −9 ). *Student's t -test versus WT P

    Article Snippet: Although our initial testing was performed using individual ER-RNAs in isolation, we used conditions designed to minimize off-target cDNA synthesis when later applied to total cellular RNA.

    Techniques: Dot Blot, Transgenic Assay, Mouse Assay, Allele-specific Oligonucleotide, Negative Control, Quantitative RT-PCR, Reverse Transcription Polymerase Chain Reaction

    piggyBac transposase bioavailability. ( A ) Half-life of the PB mRNA . Cells were transfected with 187.5 ng of PB mRNA. Total RNA was extracted at indicated times, reverse transcribed and subjected to qPCR. 18S RNA served as an internal standard to normalize the data. ( B ) Persistence of PB pDNA after transfection . Cells were transfected with 187.5 ng of PB pDNA and plasmid rescue was performed at 0 to 20 days. Ampicillin-resistant colonies were selected to evaluate the persistence of the plasmid. ( C ) Half-life of the PB transposase (V5PB Tp) . Cells were transfected with 187.5 ng of PB mRNA, incubated 18 h to reach the peak of transposase expression and treated with cycloheximide (t0=100). Total protein extraction was done at the indicated times from t0. The transposase half-life was determined by Western-Blot and quantification was normalized to the endogenous actin protein.

    Journal: PLoS ONE

    Article Title: Optimization of the piggyBac Transposon Using mRNA and Insulators: Toward a More Reliable Gene Delivery System

    doi: 10.1371/journal.pone.0082559

    Figure Lengend Snippet: piggyBac transposase bioavailability. ( A ) Half-life of the PB mRNA . Cells were transfected with 187.5 ng of PB mRNA. Total RNA was extracted at indicated times, reverse transcribed and subjected to qPCR. 18S RNA served as an internal standard to normalize the data. ( B ) Persistence of PB pDNA after transfection . Cells were transfected with 187.5 ng of PB pDNA and plasmid rescue was performed at 0 to 20 days. Ampicillin-resistant colonies were selected to evaluate the persistence of the plasmid. ( C ) Half-life of the PB transposase (V5PB Tp) . Cells were transfected with 187.5 ng of PB mRNA, incubated 18 h to reach the peak of transposase expression and treated with cycloheximide (t0=100). Total protein extraction was done at the indicated times from t0. The transposase half-life was determined by Western-Blot and quantification was normalized to the endogenous actin protein.

    Article Snippet: PB mRNA Cells were transfected with 187.5 ng of V5PB mRNA and total cellular RNA was extracted at different times using the Nucleospin RNA II kit (Macherey-Nagel, Dueren, Germany), reverse transcribed using the RevertAid First Strand cDNA Synthesis kit (Roche Applied Science, Penzberg, Germany), and subjected to real-time quantitative analysis on a MiniOpticon Real-Time PCR System instrument and software (Bio-Rad, Hercules CA, USA) with MesaGreen qPCR Master Mix Plus for SYBR Assay NoRox kit (Eurogentec, Seraing, Belgium).

    Techniques: Transfection, Real-time Polymerase Chain Reaction, Plasmid Preparation, Incubation, Expressing, Protein Extraction, Western Blot

    Effects of I3C on the expression of cell-cycle and apoptosis regulatory proteins . Western blot analysis of HUT-102 and TL-OmI cells treated with I3C. (A) Cells were treated with I3C (100 μM) for the indicated periods. (B) Cells were treated with I3C at the indicated concentrations for 48 h. Total cellular proteins (20 μg per lane) were separated on SDS-polyacrylamide gels and transferred to the membrane. Protein levels were detected by Western blotting with antibodies directed against each protein. (C) Total RNA was extracted from HUT-102 and TL-OmI cells following treatment with I3C (100 μM) for 12 or 24 h. The mRNA expression of Tax and HBZ was analyzed by RT-PCR analysis. β-actin served as an internal control in the RT-PCR procedure. Representative data of three experiments with similar results.

    Journal: Retrovirology

    Article Title: Anti-adult T-cell leukemia/lymphoma effects of indole-3-carbinol

    doi: 10.1186/1742-4690-6-7

    Figure Lengend Snippet: Effects of I3C on the expression of cell-cycle and apoptosis regulatory proteins . Western blot analysis of HUT-102 and TL-OmI cells treated with I3C. (A) Cells were treated with I3C (100 μM) for the indicated periods. (B) Cells were treated with I3C at the indicated concentrations for 48 h. Total cellular proteins (20 μg per lane) were separated on SDS-polyacrylamide gels and transferred to the membrane. Protein levels were detected by Western blotting with antibodies directed against each protein. (C) Total RNA was extracted from HUT-102 and TL-OmI cells following treatment with I3C (100 μM) for 12 or 24 h. The mRNA expression of Tax and HBZ was analyzed by RT-PCR analysis. β-actin served as an internal control in the RT-PCR procedure. Representative data of three experiments with similar results.

    Article Snippet: First-strand cDNA was synthesized from 5 μg total cellular RNA using an RNA PCR kit (Takara Bio, Otsu, Japan) with random primers.

    Techniques: Expressing, Western Blot, Reverse Transcription Polymerase Chain Reaction

    Homologous rat sequences can partially substitute for the intronic enhancer but not the silencer in intron 10 of human gene. Schematic diagram of human IR minigene chimeras with rat sequence substitutions is shown on left. White boxes represent human exons 10 and 11 separated by intron 10. Dotted lines indicate deletions in the intervening intron. Gray boxes indicate the homologous sequence from the rat Insr gene. Numbers below the introns indicate sizes of deletions. Numbers above the intron and gray boxes indicate length of human or rat sequence. The positions of the human intronic splicing enhancer ( ISE ) or silencer ( ISS ) are indicated in hIRB. These minigenes were transfected into HepG2 cells. Total RNA was isolated 48 h post-transfection and was subjected to RT-PCR analysis using primers specific to the transfected IR minigene mRNA. The mean ± S.E. for percent exon 11 inclusion is shown as a bar graph on the right . Results are derived from at least three independent experiments. Statistical significance was analyzed by ANOVA. Bars with the same letter are not significantly different ( p

    Journal: The Journal of Biological Chemistry

    Article Title: Muscleblind-like 1 (Mbnl1) Promotes Insulin Receptor Exon 11 Inclusion via Binding to a Downstream Evolutionarily Conserved Intronic Enhancer *

    doi: 10.1074/jbc.M109.095224

    Figure Lengend Snippet: Homologous rat sequences can partially substitute for the intronic enhancer but not the silencer in intron 10 of human gene. Schematic diagram of human IR minigene chimeras with rat sequence substitutions is shown on left. White boxes represent human exons 10 and 11 separated by intron 10. Dotted lines indicate deletions in the intervening intron. Gray boxes indicate the homologous sequence from the rat Insr gene. Numbers below the introns indicate sizes of deletions. Numbers above the intron and gray boxes indicate length of human or rat sequence. The positions of the human intronic splicing enhancer ( ISE ) or silencer ( ISS ) are indicated in hIRB. These minigenes were transfected into HepG2 cells. Total RNA was isolated 48 h post-transfection and was subjected to RT-PCR analysis using primers specific to the transfected IR minigene mRNA. The mean ± S.E. for percent exon 11 inclusion is shown as a bar graph on the right . Results are derived from at least three independent experiments. Statistical significance was analyzed by ANOVA. Bars with the same letter are not significantly different ( p

    Article Snippet: Cells were harvested 48 h after transfection, and total cellular RNA was prepared using RNAzol B (Tel-Test Inc., Friendswood, TX) following the manufacturer's directions and precipitated twice.

    Techniques: Sequencing, Transfection, Isolation, Reverse Transcription Polymerase Chain Reaction, Derivative Assay

    Detection of SHFV RNA by RT-PCR in baboon sera. RNA isolated from 100 μl of baboon serum is analyzed by one-step RT-PCR. Pairs of primers are designed to amplify (A) nucleocapsid or (B) helicase regions of the SHFV genome. RNA extracted from an

    Journal: Virology

    Article Title: A simian hemorrhagic fever virus isolate from persistently infected baboons efficiently induces hemorrhagic fever disease in Japanese macaques

    doi: 10.1016/j.virol.2014.10.018

    Figure Lengend Snippet: Detection of SHFV RNA by RT-PCR in baboon sera. RNA isolated from 100 μl of baboon serum is analyzed by one-step RT-PCR. Pairs of primers are designed to amplify (A) nucleocapsid or (B) helicase regions of the SHFV genome. RNA extracted from an

    Article Snippet: Total cellular RNA from SHFV-infected PBMCs was isolated with TRI Reagent (Molecular Research Center, Inc.) according to manufacturer’s protocol.

    Techniques: Reverse Transcription Polymerase Chain Reaction, Isolation

    The Inflammatory Response Depends on the RNase Activity of IRE1α and on RIG-I, a Cytosolic Sensor of RNA Fragments

    Journal: Cell host & microbe

    Article Title: The Unfolded Protein Response Element IRE1? Senses Bacterial Proteins Invading the ER to Activate RIG-I and Innate Immune Signaling

    doi: 10.1016/j.chom.2013.03.011

    Figure Lengend Snippet: The Inflammatory Response Depends on the RNase Activity of IRE1α and on RIG-I, a Cytosolic Sensor of RNA Fragments

    Article Snippet: Total cellular RNA from IRE1α/β-deleted cells was collected and harvested by solvent extraction with RNAstat60, according to the manufacturer’s instructions (AMSbio).

    Techniques: Activity Assay