Get high multiplex performance in an American-made product. Excellent sensitivity with CDC-recommended primers. All enzymes are synthesized in the United States. Fast turn-around times for customization that fits your workflow and format.
Compatible with CDC recommended RNA extraction kits and qPCR instruments.
Multiplex Detection of
This multiplex RT-qPCR assay detects two regions on the SARS-CoV-2 nucleocapsid (N) gene, N1 and N2, on two different channels. The assay uses human RNase P as the sample control gene on the third channel
Two Protocols – Same Kit
Direct: No RNA Extraction (LDT), straight from VTM/UTM*
Extracted: Use CDC Recommended RNA Extraction
2 regions of SARS-CoV-2 virus N nucleocapsid protein gene plus control region of RNaseP gene
100ng to 1pg using CDC recommended extraction protocols
AzureSeq was determined to be compatible with RNA extraction workflow at concentrations at or near LoD. Pooled negative OP swabs were spiked with genomic RNA. Input volume for extraction was 250 μl and elution volume was 80 μl. A 5 μl aliquot of the eluate was added to the PCR reaction.
Detection on nCoV2 N1 and N2 genes: 50 copies of genomic RNA added in a 20 μl reaction. Reaction set-up and RT-qPCR protocol were in accordance with CDC guidance.
Negative nasopharyngeal swabs were spiked with heat-killed SARS-CoV-2 virus at the labeled concentration in copies/uL in starting sample media (saline). Samples were heated at 95C for 5 min, then 5uL of heat-inactivated sample was added to reactions containing 1X AzureSeq CoV-2 reagents. The final reaction volumes were 20ul. Samples were run on a BioRad CFX96 using default ramp rates.
The LoD for the AzureSeq One-Step reagents for genomic RNA on the BioRad CFX96 Touch Real-time Detection System has been determined to be 5 copies per 20-μL reaction (or 0.25 copies/μL) for N1 and 10 copies per 20-μL reaction (or 0.5 copies/μL) for N2. Genomic RNA was spiked directly into PCR reactions and then analyzed on the BioRad CFX96 Touch Real- Time Detection System.
The LoD for the AzureSeq One-Step reagents for genomic RNA on the Applied Biosystems QuantStudio 3 Real-time PCR System has been determined to be 10 copies per 20-μL reaction (or 0.5 copies/μL) for both N1 and N2 targets. Genomic RNA was spiked directly into PCR reactions and then analyzed on the Quant-Studio Real-Time PCR System.
20 ul Reactions
In Solution: InhibiTaq Plus, Hot Start Mastermix, RTScript, SARS CoV-2 and RNaseP primers and probes, DTT, UDG, RNase Inhibitor
AzureSeq Direct Custom
For Research Use Only. CLIA labs must file an LDT or EUA.
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