The viral RNA was extracted from mice fecal samples using the NucleoSpin RNA Virus (Macherey-Nagel) kit following the manufacturer’s instructions. The primers and probe sequences utilized for RT-qPCR have previously been described [18 (link)]. The amplification of RNA samples was performed with one-step TaqMan RT-qPCR using the RNA UltraSense One-Step quantitative system (Thermo Fisher Scientific, Waltham, MA, USA). The standard curve was generated by serial end-point dilution, amplifying 10-fold dilutions of the quantified plasmid containing the HuNoV target sequence by RT-qPCR in triplicates. The plasmid utilized to quantify the viral load was custom synthesized by GenArt (Thermo Fisher Scientific) containing the NoV sequence previously described [18 (link)].
Rna ultrasense one step quantitative system
Manufactured by Thermo Fisher Scientific
Sourced in United States, Spain
The RNA UltraSense One-Step quantitative system is a laboratory equipment product designed for the quantitative detection of RNA. It provides a one-step approach for the reverse transcription and real-time PCR amplification of RNA samples.
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Lab products found in correlation
2 protocols using rna ultrasense one step quantitative system
1
Quantifying Human Norovirus in Mice Feces
2
RNA Extraction and RT-qPCR for Murine Stool Samples
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We extracted RNA from mice stool samples using the NucleoSpin RNA Virus (Macherey-Nagel, Duren, Germany) kit following the manufacturer’s instructions. The primers and probe sequences utilized for RT-qPCR have previously been described [15 (link)]. Amplification of RNA samples was performed with one-step TaqMan RT-qPCR using the RNA UltraSense One-Step quantitative system (Thermo Fisher Scientific, Madrid, Spain). The standard curve was generated by serial end-point dilution, amplifying 10-fold dilutions of the quantified plasmid containing the RV target sequence by RT-qPCR in triplicates.
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