Selected article for: "final concentration and PCR kit"
Author: Devon Chandler-Brown; Anna M. Bueno; Oguzhan Atay; David S. Tsao
Title: A Highly Scalable and Rapidly Deployable RNA Extraction-Free COVID-19 Assay by Quantitative Sanger Sequencing
  • Document date: 2020_4_10
    • ID: eui41zyg_36
    Snippet: The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/2020.04.07.029199 doi: bioRxiv preprint qSanger Amplification: Reverse transcription and amplification for figure 2 was performed using OneTaq One-Step RT-PCR Kit from NEB (cat. # E5315S). Both the OneTaq One-Step RT-PCR Kit and Luna Universal One-step RT-qPCR Kit (cat. # E3005E) were used for figure 3. Figure 4 was performed exclusively with the.....
    Document: The copyright holder for this preprint (which was not peer-reviewed) is the . https://doi.org/10.1101/2020.04.07.029199 doi: bioRxiv preprint qSanger Amplification: Reverse transcription and amplification for figure 2 was performed using OneTaq One-Step RT-PCR Kit from NEB (cat. # E5315S). Both the OneTaq One-Step RT-PCR Kit and Luna Universal One-step RT-qPCR Kit (cat. # E3005E) were used for figure 3. Figure 4 was performed exclusively with the Luna Universal One-step RT-qPCR Kit. Buffer and enzyme were used according to manufacturer recommendations for 100 µL total volume. All reactions contained Tween-20 at a final concentration of 1% v/v, 500 nM final concentration of each amplification primer, and 100 GCE of synthetic dsDNA spike-in molecules. Synthetic RNA samples were added at 2 µL/reaction and synthetic sample was added to achieve the appropriate number of viral particles. Thermocycling was performed using an Applied Biosystems Veriti Thermal Cycler with the following cycling programs:

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