goscript™ rt mix for 1-step rt-qpcr (50×)  (Promega)

Journal: medRxiv

Article Title: A handheld point-of-care system for rapid detection of SARS-CoV-2 in under 20 minutes

doi: 10.1101/2020.06.29.20142349

Figure Lengend Snippet: SARS-CoV-2 diagnostics workflow. (A) Sample collection and preparation illustrating nasopharyngeal swab and RNA extraction. (B) Nucleic acid amplification methods for SARS-CoV-2 RNA detection used in this study (RT-qPCR, RT-qLAMP and RT-eLAMP). Thermal profiles are illustrated for assays comparison. (C) Point-of-care diagnostics by RT-eLAMP showing the proposed handheld LoC platform including the microfluidic cartridge with control and sample inlets, and the smartphone-enabled application for geo-localization and real-time visualization of results.

Article Snippet: Each mix contained the following: 10 μL of 2× GoTaq® qPCR master mix, 0.4 μL of 50× GoScript™ RT mix for 1-Step RT-qPCR, 1.2 μL of 16.6× N1/RNase P assay primer mix by the CDC (Forward Primer (20 μM), Reverse Primer (20 μM), Probe (5 μM)), 4 μL of extracted RNA and enough nuclease-free water (GoTaq® 1-Step RT-qPCR kit, Promega) to bring the volume to 20 μL.

Techniques: RNA Extraction, Amplification, RNA Detection, Quantitative RT-PCR

Journal: medRxiv

Article Title: A handheld point-of-care system for rapid detection of SARS-CoV-2 in under 20 minutes

doi: 10.1101/2020.06.29.20142349

Figure Lengend Snippet:

Article Snippet: Each mix contained the following: 10 μL of 2× GoTaq® qPCR master mix, 0.4 μL of 50× GoScript™ RT mix for 1-Step RT-qPCR, 1.2 μL of 16.6× N1/RNase P assay primer mix by the CDC (Forward Primer (20 μM), Reverse Primer (20 μM), Probe (5 μM)), 4 μL of extracted RNA and enough nuclease-free water (GoTaq® 1-Step RT-qPCR kit, Promega) to bring the volume to 20 μL.

Techniques:

Journal: medRxiv

Article Title: A handheld point-of-care system for rapid detection of SARS-CoV-2 in under 20 minutes

doi: 10.1101/2020.06.29.20142349

Figure Lengend Snippet: Clinical validation using RT-qPCR, RT-qLAMP and RT-eLAMP. (A) Correlation between RT-qPCR and RT-qLAMP based on the estimated sample concentration (copies/μL of eluted sample volume), n = 115. (B) Correlation between RNase P and viral RNA concentration of the clinical samples (copies/µL of eluted sample volume) to indicate quality of extraction across all the cohort, n = 127. (C) Boxplot distribution of RNase P concentration (copies/µL of eluted sample volume) across negative (n = 56) and positive (n = 127) clinical samples by RT-qPCR. Calculated p-value between both groups was below 0.05 (p-value = 8.09×10 −6 ). (D) Boxplot distribution of TTP for the RT-qLAMP and RT-eLAMP demonstrating the performance of the LoC platform. qPCR. Calculated p-value between both groups is higher than 0.05 (p-value = 0.32). (E) Example overview of data processing steps for sample #179 to extract amplification curves from sample and control wells on the LoC platform during RT-eLAMP. The spatial image illustrates the microchip ISFET sensing array output (4,368 sensors, 2 × 4 mm) on the single-use cartridge where the averaged sensor signals in each well demonstrate respectively amplification with a TTP of 10.63 min (sample) and no amplification (control).

Article Snippet: Each mix contained the following: 10 μL of 2× GoTaq® qPCR master mix, 0.4 μL of 50× GoScript™ RT mix for 1-Step RT-qPCR, 1.2 μL of 16.6× N1/RNase P assay primer mix by the CDC (Forward Primer (20 μM), Reverse Primer (20 μM), Probe (5 μM)), 4 μL of extracted RNA and enough nuclease-free water (GoTaq® 1-Step RT-qPCR kit, Promega) to bring the volume to 20 μL.

Techniques: Quantitative RT-PCR, Concentration Assay, Amplification, MicroChIP Assay