Description
General description
The Viral RNA Extraction Buffer has been optimized to provide rapid, room-temperature lysis of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) viral particles in saliva. The buffer comes as a 4x working solution that can be mixed with virus-containing samples in a qRT-PCR plate. Upon 5 minute incubation at room temperature, viral particles are lysed and RNA is stabilized such that samples are ready for direct analysis via RT-qPCR with no additional cleanup step. Removal of precipitates in saliva via centrifugation prior to testing is recommended for optimal sensitivity.
Application
The Viral RNA Extraction Buffer is broadly compatible with most nucleic acid detection methods and does not reduce signal when used at working concentration in qRT-PCR. qRT-PCR compatibility has been demonstrated in a 4-fluor multiplex setting consisting of Cy5, TxRed, FAM, and HEX.
RT-qPCR compatibility has been verified with several one-step RT-qPCR kits:
Quantitative RT-PCR ReadyMix™ (Sigma-Aldrich, catalog QR0200)
KAPA PROBE FAST One-Step Universal (Sigma-Aldrich, catalog KK4752)
KiCqStart® One-Step Probe RT-qPCR ReadyMix™ (Sigma-Aldrich, catalog KCQS07)
TaqPathTM 1-Step Multiplex Master Mix (Thermo Fisher, catalog A28525)
The Viral RNA Extraction Buffer is also compatible with colorimetric LAMP assays.
Features and Benefits
Extracted viral RNA can be used for detection without cleanup
Directly compatible with quantitative reverse transcription polymerase chain reaction (qRT-PCR) and loop mediated isothermal amplification(LAMP) assays
Ideal for enveloped RNA viruses in saliva, saline, Amies medium, and viral transport medium
Highly sensitive and faster than other leading methods, lysis by Proteinase K and heating
Legal Information
KiCqStart is a registered trademark of QIAGEN Beverly Inc.
ReadyMix is a trademark of Sigma-Aldrich Co. LLC
