A Fast and Accessible Method for the Isolation of RNA, DNA, and Protein to Facilitate the Detection of SARS-CoV-2
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Abstract
Management of the COVID-19 pandemic requires widespread SARS-CoV-2 testing. A main limitation for widespread SARS-CoV-2 testing is the global shortage of essential supplies, among these, RNA extraction kits. The need for commercial RNA extraction kits places a bottleneck on tests that detect SARS-CoV-2 genetic material, including PCR-based reference tests. Here we propose an alternative method we call PEARL ( P recipitation E nhanced A nalyte R etrieva L ) that addresses this limitation. PEARL uses a lysis solution that disrupts cell membranes and viral envelopes while simultaneously providing conditions suitable for alcohol-based precipitation of RNA, DNA, and proteins. PEARL is a fast, low-cost, and simple method that uses common laboratory reagents and offers comparable performance to commercial RNA extraction kits. PEARL offers an alternative method to isolate host and pathogen nucleic acids and proteins to streamline the detection of DNA and RNA viruses, including SARS-CoV-2.
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SciScore for 10.1101/2020.06.29.178384: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement not detected. Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources Primary antibodies were diluted in 0.5% BSA-TBST as follows: anti-HSP-70 (Cell Signaling Technology 4872) 1:1,000; anti-LANA/ORF73 (Advanced Biotechnologies 13-210-100) anti-HSP-70 ( Cell Signaling Technology 4872suggested: Noneanti-LANA/ORF73suggested: NoneExperimental Models: Cell Lines Sentences Resources For ZIKV infections, HeLa cells were grown to 60% confluency and then infected with ZIKV at a multiplicity of infection (MOI) of 1. HeLasuggested: None… SciScore for 10.1101/2020.06.29.178384: (What is this?)
Please note, not all rigor criteria are appropriate for all manuscripts.
Table 1: Rigor
Institutional Review Board Statement not detected. Randomization not detected. Blinding not detected. Power Analysis not detected. Sex as a biological variable not detected. Cell Line Authentication not detected. Table 2: Resources
Antibodies Sentences Resources Primary antibodies were diluted in 0.5% BSA-TBST as follows: anti-HSP-70 (Cell Signaling Technology 4872) 1:1,000; anti-LANA/ORF73 (Advanced Biotechnologies 13-210-100) anti-HSP-70 ( Cell Signaling Technology 4872suggested: Noneanti-LANA/ORF73suggested: NoneExperimental Models: Cell Lines Sentences Resources For ZIKV infections, HeLa cells were grown to 60% confluency and then infected with ZIKV at a multiplicity of infection (MOI) of 1. HeLasuggested: NoneSoftware and Algorithms Sentences Resources Image post-processing was carried out in Photoshop CC (Adobe) using automatic contrast. Photoshop CCsuggested: (Adobe Photoshop, RRID:SCR_014199)Results from OddPub: We did not detect open data. We also did not detect open code. Researchers are encouraged to share open data when possible (see Nature blog).
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.
- No funding statement was detected.
- No protocol registration statement was detected.
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