The Applied Biosystems TaqMan SARS-CoV-2 Mutation Panel is a scalable solution for your SARS-CoV-2 mutation-screening needs. This panel of assays lets you run a variable number of samples, from just a few to dozens and hundreds, to monitor for one or many SARS-CoV-2 mutations, and it can do this on your current real-time PCR instrument. The secret is, our process combines gold-standard Applied Biosystems TaqMan SNP genotyping technology with a one-step real-time PCR reaction to detect mutations in a sample quickly and efficiently, directly from the RNA. But how does it work?
Starting with RNA isolated from SARS-CoV-2-positive samples, reverse transcription turns that RNA into much more stable cDNA. This is necessary for the rest of the reactions to proceed without damaging the sample, and our system combines this step into the rest of the reactions instead of requiring that it be done separately.
Once the cDNA is ready, PCR amplification begins. Our master mix contains a DNA polymerase formulated for high-temperature applications like PCR, forward and reverse primers, a FAM-labeled TaqMan probe for the mutant sequence of interest, and a VIC-labeled TaqMan probe for the corresponding wild-type sequence. The thermal cycler denatures the cDNA templates, separating their strands so that the primers can anneal to their target sequences in the cDNA and the probes can anneal between them. During polymerization, DNA polymerase turns these primers and probes into whole complementary DNA strands, doubling the amount of DNA in the reaction and, importantly, separating the probes from their built-in quenchers. Once the quenchers are separated, the probes’ fluorescence increases, and it is this difference that the real-time PCR system collects as data. Our use of probes optimized for their target sequences removes the hazard of off-target activation and makes sure that the reading clearly indicates mutant or wild-type presence in the sample, as appropriate.
After data collection, you can analyze your data with Applied Biosystems Design and Analysis Software 2.5 or later, using the Genotyping Analysis Module. The results are shown as scatter plots, with samples that contain the mutant allele clustered away from those that are wild-type. Patterns in which samples cluster with each other will illuminate your research question and increase knowledge of SARS-CoV-2’s mutation patterns.
To learn more about the TaqMan SARS-CoV-2 Mutation Panel, visit thermofisher.com/mutationpanel.
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