Selected article for: "positive control and rnase free water"
Author: Xianding Deng; Wei Gu; Scot Federman; Louis Du Plessis; Oliver Pybus; Nuno Faria; Candace Wang; Guixia Yu; Chao-Yang Pan; Hugo Guevara; Alicia Sotomayor-Gonzalez; Kelsey Zorn; Allan Gopez; Venice Servellita; Elaine Hsu; Steve Miller; Trevor Bedford; Alexander Greninger; Pavitra Roychoudhury; Michael Famulare; Helen Y Chu; Jay Shendure; Lea Starita; Catie Anderson; Karthik Gangavarapu; Mark Zeller; Emily Spencer; Kristian Andersen; Duncan MacCannell; Suxiang Tong; Gregory Armstrong; Clinton Paden; Yan Li; Ying Zhang; Scott Morrow; Matthew Willis; Bela Matyas; Sundari Mase; Olivia Kasirye; Maggie Park; Curtis Chan; Alexander Yu; Shua Chai; Elsa Villarino; Brandon Bonin; Debra Wadford; Charles Y Chiu
Title: A Genomic Survey of SARS-CoV-2 Reveals Multiple Introductions into Northern California without a Predominant Lineage
  • Document date: 2020_3_30
    • ID: cbc98t7x_27
    Snippet: is the (which was not peer-reviewed) The copyright holder for this preprint (Qiagen, Hillden, Germany) . At the California Department of Public Health, samples were extracted using Qiagen DSP Viral RNA mini kit with carrier RNA added (Qiagen). The one-step RT-PCR used the 2019-nCoV CDC qPCR Probe Assay targets N1 and N2, and the 2019-nCoV N Positive Control plasmid (Integrated DNA Technologies Inc. IDT, Coralville, USA) as a positive control. The.....
    Document: is the (which was not peer-reviewed) The copyright holder for this preprint (Qiagen, Hillden, Germany) . At the California Department of Public Health, samples were extracted using Qiagen DSP Viral RNA mini kit with carrier RNA added (Qiagen). The one-step RT-PCR used the 2019-nCoV CDC qPCR Probe Assay targets N1 and N2, and the 2019-nCoV N Positive Control plasmid (Integrated DNA Technologies Inc. IDT, Coralville, USA) as a positive control. The RNA extracts were added to an amplification reaction mix (4X TaqPath 1-step RT-PCR master mix (Life Technologies, Carlsbad, USA), 1.5 ul primer (N1, N2, or RNase P), 12.5ul nuclease-free water, and 1ul RNA) and one-step RT-PCR was performed (25ºC for 2 mins, reverse transcription at 50ºC for 15 mins, 95ºC for 2 mins, followed by 45 cycles PCR with denaturing at 95ºC for 3s and extension at 55ºC for 30s). The cut-off for a confirmed positive sample was determined to be at a Ct value of 40 cycles, with both targets of N1 and N2 necessarily being classified as positive by quantitative RT-PCR, except for a positive RNase P as an internal quality control.

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