Massively scaled-up testing for SARS-CoV-2 RNA via next-generation sequencing of pooled and barcoded nasal and saliva samples

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Abstract

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  1. SciScore for 10.1101/2020.08.04.20167874: (What is this?)

    Please note, not all rigor criteria are appropriate for all manuscripts.

    Table 1: Rigor

    Institutional Review Board Statementnot detected.
    Randomizationnot detected.
    Blindingnot detected.
    Power Analysisnot detected.
    Sex as a biological variablenot detected.
    Cell Line Authenticationnot detected.

    Table 2: Resources

    Experimental Models: Cell Lines
    SentencesResources
    RT PCR (FP_1, R) and a second round of PCR (FP_2, R) was performed on HEK293T lysate for construction of an in vitro RPP30 standard DNA template.
    HEK293T
    suggested: None
    Software and Algorithms
    SentencesResources
    We also tested 1x Tween with Qiagen Protease and RNA Secure (ThermoFisher), which also works but resulted in more sample-to-sample variability and required additional incubation steps.
    ThermoFisher
    suggested: (ThermoFisher; SL 8; Centrifuge, RRID:SCR_020809)
    Purified RNA was quantified and library quality was assessed using the Agilent TapeStation.
    Agilent TapeStation
    suggested: (Agilent TapeStation Laptop, RRID:SCR_019547)
    On the NextSeq550 and MiniSeq, the post-run wash was performed automatically by the instruments, and no human intervention was required.
    MiniSeq
    suggested: None
    For this application, the MiSeq and MiniSeq (Rapid Run Kit) require 2 custom sequencing primer mixes, the Read1 primer mix and the i7 primer mix.
    MiSeq
    suggested: (A5-miseq, RRID:SCR_012148)
    Analysis: The bioinformatic analysis consists of standard conversion of BCL files into FASTQ sequencing files using Illumina’s bcl2fastq software (v2.20.0.422).
    Illumina’s
    suggested: None
    bcl2fastq
    suggested: (bcl2fastq , RRID:SCR_015058)
    In this analysis we make use of a few custom scripts written in R that rely on the ShortRead 30 and stringdist 31 packages for processing fastq files and calculating hamming distances between observed and expected amplicons and indices.
    ShortRead
    suggested: (ShortRead, RRID:SCR_006813)

    Results from OddPub: Thank you for sharing your code and data.


    Results from LimitationRecognizer: An explicit section about the limitations of the techniques employed in this study was not found. We encourage authors to address study limitations.

    Results from TrialIdentifier: No clinical trial numbers were referenced.


    Results from Barzooka: We did not find any issues relating to the usage of bar graphs.


    Results from JetFighter: We did not find any issues relating to colormaps.


    Results from rtransparent:
    • Thank you for including a conflict of interest statement. Authors are encouraged to include this statement when submitting to a journal.
    • Thank you for including a funding statement. Authors are encouraged to include this statement when submitting to a journal.
    • No protocol registration statement was detected.

    About SciScore

    SciScore is an automated tool that is designed to assist expert reviewers by finding and presenting formulaic information scattered throughout a paper in a standard, easy to digest format. SciScore checks for the presence and correctness of RRIDs (research resource identifiers), and for rigor criteria such as sex and investigator blinding. For details on the theoretical underpinning of rigor criteria and the tools shown here, including references cited, please follow this link.