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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 18 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Average 90 stars, based on 18 article reviews
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rgtp - by Bioz Stars, 2020-08
90/100 stars

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1) Product Images from "Purification and Biochemical Characterisation of Rabbit Calicivirus RNA-Dependent RNA Polymerases and Identification of Non-Nucleoside Inhibitors"

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

Journal: Viruses

doi: 10.3390/v8040100

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.
Figure Legend 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.

Techniques Used: 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.
Figure Legend 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.

Techniques Used: 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.
Figure Legend 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.

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

2) Product Images from "Purification and Biochemical Characterisation of Rabbit Calicivirus RNA-Dependent RNA Polymerases and Identification of Non-Nucleoside Inhibitors"

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

Journal: Viruses

doi: 10.3390/v8040100

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.
Figure Legend 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.

Techniques Used: 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.
Figure Legend 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.

Techniques Used: 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.
Figure Legend 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.

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

Related Articles

Polymerase Chain Reaction:

Article Title: Defining a novel leptin–melanocortin–kisspeptin pathway involved in the metabolic control of puberty
Article Snippet: .. For the generation of the antisense Kiss1 riboprobe, the product of the PCR was used as template for the transcription reaction, as follows (final volume of 20 μl): 250 Ci [33P]-UTP (Perkin Elmer, Massachusetts, USA), 0.5 μg of template, 2 μl 3NTPs (5 mM rATP, rCTP and rGTP), 1 μl RNasin Ribonuclease Inhibitor (Promega), 4 μl transcription buffer, and 2 μl T7 RNA polymerase (Promega). .. After 120 min of incubation at 37 °C, another 1 μl of T7 RNA was added to the mix, and the reaction was maintained for an additional 60 min at 37 °C.

Incubation:

Article Title: The extruded non-template strand determines the architecture of R-loops
Article Snippet: .. In vitro transcription of R-loops 1875 ng of circular plasmids were incubated at 37°C with 50 U of T3 RNA Polymerase (Promega) in a transcription buffer (40 mM Tris–HCl pH 7.9, 6 mM MgCl2 , 2 mM spermidine, 10 mM NaCl, 20 mM DTT, 0,05% Tween-20) containing 0,25 mM of rATP, rCTP, rUTP and rGTP (Promega). ..

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Article Snippet: .. The 100-μl reaction mixture, containing 20 μl of 5× T7 buffer; 7.5 mM (each) rATP, rCTP, rGTP, and rUTP; 2 μg of DNA; and 10 μl of enzyme mixture was incubated at 37°C for 2 h and then treated with 10 U of RNase-free DNase (Promega) for 30 min at 37°C. ..

Article Title: A Nucleus-Encoded Chloroplast Phosphoprotein Governs Expression of the Photosystem I Subunit PsaC in Chlamydomonas reinhardtii
Article Snippet: .. Then, 40 μL 4× run-on buffer (100 mM HEPES, pH 7.5, 1 M sucrose, 120 mM MgCl2 , 30 mM DTT, and 100 mM NaF) was mixed with 5 μL each of 10 mM rATP, rGTP, and rCTP, 5 μL RNasin (Promega), 20 μL 32 P-rUTP (200 µCi, 5 µM final), and 80 μL of cell pellet (defrosted in a water bath at 20°C).The reaction mixture was incubated at 26°C for 5 or 15 min. Total RNA was immediately extracted with TRI Reagent (Sigma-Aldrich), precipitated with isopropanol, and resuspended in TE (10 mM Tris and 1 mM EDTA, pH 8) prior to separation on a Sephadex G50 column. .. The RNA fractions were then used for hybridization.

In Vitro:

Article Title: The extruded non-template strand determines the architecture of R-loops
Article Snippet: .. In vitro transcription of R-loops 1875 ng of circular plasmids were incubated at 37°C with 50 U of T3 RNA Polymerase (Promega) in a transcription buffer (40 mM Tris–HCl pH 7.9, 6 mM MgCl2 , 2 mM spermidine, 10 mM NaCl, 20 mM DTT, 0,05% Tween-20) containing 0,25 mM of rATP, rCTP, rUTP and rGTP (Promega). ..

Plasmid Preparation:

Article Title: TALEN/CRISPR-mediated engineering of a promoterless anti-viral RNAi hairpin into an endogenous miRNA locus
Article Snippet: .. For HCV transcripts and use with SP6 polymerase, the mix contained 80 mM HEPES (pH 7.5), 16 mM MgCl2 , 2 mM spermidine, 40 mM DTT, 3.125 mM each of rATP, rCTP and rUTP, 1.5625 mM of rGTP, 1 mM m7 G(5′)ppp(5′)G RNA cap structure analog, 1 U/μl of RNasin, 0.1 μg/μl of plasmid DNA and 0.6 U/μl of SP6 RNA polymerase (Promega). .. After a 2 h incubation at 37°C (HCV) or 40°C (Dengue virus), 0.3 U of T7 RNA polymerase or 0.4 U of SP6 RNA polymerase, respectively, were added per μl of reaction mixture, and the reaction mixture was incubated overnight.

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    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 18 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rgtp/product/Promega
    Average 90 stars, based on 18 article reviews
    Price from $9.99 to $1999.99
    rgtp - by Bioz Stars, 2020-08
    90/100 stars
      Buy from Supplier

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    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