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  • 90
    Promega rgtp
    Impact of MnCl 2 , pH and temperature on calicivirus RdRp activity. Purified recombinant RdRps (40 ng of RHDV and RCV RdRps and 200 ng of NoV RdRp) were used to generate dsRNA with poly(C) <t>RNA</t> (20 ng/μL) as template and <t>rGTP</t> (0.5 mM) as substrate. Following a 15-min incubation in the presence of different ( a ) MnCl 2 concentrations; ( b ) pH levels; or ( c ) temperatures, reactions were stopped with EDTA at a final concentration 5 mM and dsRNA was quantified using the PicoGreen reagent. Unless indicated otherwise, reactions were incubated at 30 °C. RHDV RdRp activity is shown in red as triangles, RCV RdRp activity is shown in blue as squares and NoV RdRp activity is shown in black as circles. Averages of relative fluorescence levels were calculated and plotted with standard deviations. The results were generated from two ( a ) or three ( b and c ) independent experiments with triplicate reactions for each measurement point.
    Rgtp, supplied by Promega, used in various techniques. Bioz Stars score: 90/100, based on 163 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rgtp/product/Promega
    Average 90 stars, based on 163 article reviews
    Price from $9.99 to $1999.99
    rgtp - by Bioz Stars, 2020-08
    90/100 stars
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    90
    Promega rntp
    Impact of MnCl 2 , pH and temperature on calicivirus RdRp activity. Purified recombinant RdRps (40 ng of RHDV and RCV RdRps and 200 ng of NoV RdRp) were used to generate dsRNA with poly(C) <t>RNA</t> (20 ng/μL) as template and <t>rGTP</t> (0.5 mM) as substrate. Following a 15-min incubation in the presence of different ( a ) MnCl 2 concentrations; ( b ) pH levels; or ( c ) temperatures, reactions were stopped with EDTA at a final concentration 5 mM and dsRNA was quantified using the PicoGreen reagent. Unless indicated otherwise, reactions were incubated at 30 °C. RHDV RdRp activity is shown in red as triangles, RCV RdRp activity is shown in blue as squares and NoV RdRp activity is shown in black as circles. Averages of relative fluorescence levels were calculated and plotted with standard deviations. The results were generated from two ( a ) or three ( b and c ) independent experiments with triplicate reactions for each measurement point.
    Rntp, supplied by Promega, used in various techniques. Bioz Stars score: 90/100, based on 149 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rntp/product/Promega
    Average 90 stars, based on 149 article reviews
    Price from $9.99 to $1999.99
    rntp - by Bioz Stars, 2020-08
    90/100 stars
      Buy from Supplier

    89
    Promega ntp mix
    Impact of MnCl 2 , pH and temperature on calicivirus RdRp activity. Purified recombinant RdRps (40 ng of RHDV and RCV RdRps and 200 ng of NoV RdRp) were used to generate dsRNA with poly(C) <t>RNA</t> (20 ng/μL) as template and <t>rGTP</t> (0.5 mM) as substrate. Following a 15-min incubation in the presence of different ( a ) MnCl 2 concentrations; ( b ) pH levels; or ( c ) temperatures, reactions were stopped with EDTA at a final concentration 5 mM and dsRNA was quantified using the PicoGreen reagent. Unless indicated otherwise, reactions were incubated at 30 °C. RHDV RdRp activity is shown in red as triangles, RCV RdRp activity is shown in blue as squares and NoV RdRp activity is shown in black as circles. Averages of relative fluorescence levels were calculated and plotted with standard deviations. The results were generated from two ( a ) or three ( b and c ) independent experiments with triplicate reactions for each measurement point.
    Ntp Mix, supplied by Promega, used in various techniques. Bioz Stars score: 89/100, based on 61 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/ntp mix/product/Promega
    Average 89 stars, based on 61 article reviews
    Price from $9.99 to $1999.99
    ntp mix - by Bioz Stars, 2020-08
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    95
    Promega ribonucleotide triphosphates
    Impact of MnCl 2 , pH and temperature on calicivirus RdRp activity. Purified recombinant RdRps (40 ng of RHDV and RCV RdRps and 200 ng of NoV RdRp) were used to generate dsRNA with poly(C) <t>RNA</t> (20 ng/μL) as template and <t>rGTP</t> (0.5 mM) as substrate. Following a 15-min incubation in the presence of different ( a ) MnCl 2 concentrations; ( b ) pH levels; or ( c ) temperatures, reactions were stopped with EDTA at a final concentration 5 mM and dsRNA was quantified using the PicoGreen reagent. Unless indicated otherwise, reactions were incubated at 30 °C. RHDV RdRp activity is shown in red as triangles, RCV RdRp activity is shown in blue as squares and NoV RdRp activity is shown in black as circles. Averages of relative fluorescence levels were calculated and plotted with standard deviations. The results were generated from two ( a ) or three ( b and c ) independent experiments with triplicate reactions for each measurement point.
    Ribonucleotide Triphosphates, supplied by Promega, used in various techniques. Bioz Stars score: 95/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 95 stars, based on 16 article reviews
    Price from $9.99 to $1999.99
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    85
    Promega nucleotide triphosphate mix
    Impact of MnCl 2 , pH and temperature on calicivirus RdRp activity. Purified recombinant RdRps (40 ng of RHDV and RCV RdRps and 200 ng of NoV RdRp) were used to generate dsRNA with poly(C) <t>RNA</t> (20 ng/μL) as template and <t>rGTP</t> (0.5 mM) as substrate. Following a 15-min incubation in the presence of different ( a ) MnCl 2 concentrations; ( b ) pH levels; or ( c ) temperatures, reactions were stopped with EDTA at a final concentration 5 mM and dsRNA was quantified using the PicoGreen reagent. Unless indicated otherwise, reactions were incubated at 30 °C. RHDV RdRp activity is shown in red as triangles, RCV RdRp activity is shown in blue as squares and NoV RdRp activity is shown in black as circles. Averages of relative fluorescence levels were calculated and plotted with standard deviations. The results were generated from two ( a ) or three ( b and c ) independent experiments with triplicate reactions for each measurement point.
    Nucleotide Triphosphate Mix, supplied by Promega, used in various techniques. Bioz Stars score: 85/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/nucleotide triphosphate mix/product/Promega
    Average 85 stars, based on 6 article reviews
    Price from $9.99 to $1999.99
    nucleotide triphosphate mix - by Bioz Stars, 2020-08
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    Image Search Results


    Impact of MnCl 2 , pH and temperature on calicivirus RdRp activity. Purified recombinant RdRps (40 ng of RHDV and RCV RdRps and 200 ng of NoV RdRp) were used to generate dsRNA with poly(C) RNA (20 ng/μL) as template and rGTP (0.5 mM) as substrate. Following a 15-min incubation in the presence of different ( a ) MnCl 2 concentrations; ( b ) pH levels; or ( c ) temperatures, reactions were stopped with EDTA at a final concentration 5 mM and dsRNA was quantified using the PicoGreen reagent. Unless indicated otherwise, reactions were incubated at 30 °C. RHDV RdRp activity is shown in red as triangles, RCV RdRp activity is shown in blue as squares and NoV RdRp activity is shown in black as circles. Averages of relative fluorescence levels were calculated and plotted with standard deviations. The results were generated from two ( a ) or three ( b and c ) independent experiments with triplicate reactions for each measurement point.

    Journal: Viruses

    Article Title: Purification and Biochemical Characterisation of Rabbit Calicivirus RNA-Dependent RNA Polymerases and Identification of Non-Nucleoside Inhibitors

    doi: 10.3390/v8040100

    Figure Lengend Snippet: Impact of MnCl 2 , pH and temperature on calicivirus RdRp activity. Purified recombinant RdRps (40 ng of RHDV and RCV RdRps and 200 ng of NoV RdRp) were used to generate dsRNA with poly(C) RNA (20 ng/μL) as template and rGTP (0.5 mM) as substrate. Following a 15-min incubation in the presence of different ( a ) MnCl 2 concentrations; ( b ) pH levels; or ( c ) temperatures, reactions were stopped with EDTA at a final concentration 5 mM and dsRNA was quantified using the PicoGreen reagent. Unless indicated otherwise, reactions were incubated at 30 °C. RHDV RdRp activity is shown in red as triangles, RCV RdRp activity is shown in blue as squares and NoV RdRp activity is shown in black as circles. Averages of relative fluorescence levels were calculated and plotted with standard deviations. The results were generated from two ( a ) or three ( b and c ) independent experiments with triplicate reactions for each measurement point.

    Article Snippet: A standard reaction contained 20 ng/μL poly(C) RNA (Sigma-Aldrich), 0.5 mM rGTP (Promega), 2.5 mM MnCl2 , and 5 mM DTT in 20 mM Tris-HCl (pH 7.5) buffer.

    Techniques: Activity Assay, Purification, Recombinant, Incubation, Concentration Assay, Fluorescence, Generated

    Comparison of RHDV and RCV RdRp activities. Purified recombinant RdRps were used to generate dsRNA from a poly(C) RNA (20 ng/μL) template using rGTP (0.5 mM) as substrate. Following incubation at 30 °C, reactions were stopped with 5 mM EDTA and dsRNA was quantified using the fluorescent dye PicoGreen. RHDV RdRp activity is shown in red, RCV RdRp activity is shown in blue. The results from a representative experiment are shown with average values and standard deviations from triplicate reactions for each measurement point. ( a ) The effect of RHDV and RCV RdRp concentrations on dsRNA formation over 15 min; ( b ) The synthesis of dsRNA catalysed by 20 ng of RHDV and RCV RdRps over a 1-h period. RFU—relative fluorescence units.

    Journal: Viruses

    Article Title: Purification and Biochemical Characterisation of Rabbit Calicivirus RNA-Dependent RNA Polymerases and Identification of Non-Nucleoside Inhibitors

    doi: 10.3390/v8040100

    Figure Lengend Snippet: Comparison of RHDV and RCV RdRp activities. Purified recombinant RdRps were used to generate dsRNA from a poly(C) RNA (20 ng/μL) template using rGTP (0.5 mM) as substrate. Following incubation at 30 °C, reactions were stopped with 5 mM EDTA and dsRNA was quantified using the fluorescent dye PicoGreen. RHDV RdRp activity is shown in red, RCV RdRp activity is shown in blue. The results from a representative experiment are shown with average values and standard deviations from triplicate reactions for each measurement point. ( a ) The effect of RHDV and RCV RdRp concentrations on dsRNA formation over 15 min; ( b ) The synthesis of dsRNA catalysed by 20 ng of RHDV and RCV RdRps over a 1-h period. RFU—relative fluorescence units.

    Article Snippet: A standard reaction contained 20 ng/μL poly(C) RNA (Sigma-Aldrich), 0.5 mM rGTP (Promega), 2.5 mM MnCl2 , and 5 mM DTT in 20 mM Tris-HCl (pH 7.5) buffer.

    Techniques: Purification, Recombinant, Incubation, Activity Assay, Fluorescence

    Kinetics of template and substrate utilisation by calicivirus RdRps. Purified recombinant RdRps (40 ng of RHDV and RCV RdRps and 200 ng of NoV RdRp) were used to generate dsRNA with poly(C) RNA as template and rGTP as substrate. ( a ) The kinetics of template utilisation was examined by titrating poly(C) from 0 to 40 ng/μL in the presence of 0.5 mM rGTP; ( b , c ) The kinetics of nucleotide incorporation was examined by titrating rGTP from 0 to 0.5 mM and 0 to 2.0 mM in the presence of 20 ng/μL of poly(C). Following a 15-min incubation at 30 °C, reactions were stopped with EDTA at final concentration 5 mM and dsRNA was quantified using the PicoGreen reagent. RHDV RdRp activity is shown in red as triangles, RCV RdRp activity is shown in blue as squares and NoV RdRp activity is shown in black as circles. The results from a representative experiment are shown with average values and standard deviations from triplicate reactions for each measurement point.

    Journal: Viruses

    Article Title: Purification and Biochemical Characterisation of Rabbit Calicivirus RNA-Dependent RNA Polymerases and Identification of Non-Nucleoside Inhibitors

    doi: 10.3390/v8040100

    Figure Lengend Snippet: Kinetics of template and substrate utilisation by calicivirus RdRps. Purified recombinant RdRps (40 ng of RHDV and RCV RdRps and 200 ng of NoV RdRp) were used to generate dsRNA with poly(C) RNA as template and rGTP as substrate. ( a ) The kinetics of template utilisation was examined by titrating poly(C) from 0 to 40 ng/μL in the presence of 0.5 mM rGTP; ( b , c ) The kinetics of nucleotide incorporation was examined by titrating rGTP from 0 to 0.5 mM and 0 to 2.0 mM in the presence of 20 ng/μL of poly(C). Following a 15-min incubation at 30 °C, reactions were stopped with EDTA at final concentration 5 mM and dsRNA was quantified using the PicoGreen reagent. RHDV RdRp activity is shown in red as triangles, RCV RdRp activity is shown in blue as squares and NoV RdRp activity is shown in black as circles. The results from a representative experiment are shown with average values and standard deviations from triplicate reactions for each measurement point.

    Article Snippet: A standard reaction contained 20 ng/μL poly(C) RNA (Sigma-Aldrich), 0.5 mM rGTP (Promega), 2.5 mM MnCl2 , and 5 mM DTT in 20 mM Tris-HCl (pH 7.5) buffer.

    Techniques: Purification, Recombinant, Incubation, Concentration Assay, Activity Assay