Apart from detecting specific targets like HF183, labs conduct additional simultaneous reactions to monitor the success and integrity of the overall assay. Understanding the roles of SKETA, IAC, pass/fail outcomes, and inhibition results in dPCR assists in ensuring accurate and reliable molecular analysis.
What is SKETA and IAC in dPCR?
SKETA: This primer and probe set specifically target salmon DNA. Before extraction, samples are spiked with salmon DNA to assess extraction success and identify inhibitory factors.
Internal Amplification Control (IAC): Is a crucial element in every dPCR reaction. The IAC confirms true negative outcomes and sheds light on potential extraction anomalies.
Both SKETA and IAC play critical roles in comprehending the molecular composition of the reaction and determining its success. SKETA detects salmon DNA presence, while IAC serves as an internal check for successful amplification.
What does “pass/fail” and “inhibition” result mean in dPCR?
Pass/Fail: These outcomes gauge the success and integrity of the assay beyond specific target detection like HF183.
“Inhibition” Result: When a sample lacks detection of both SKETA and IAC, it indicates potential inhibition hindering the PCR process, making interpretation challenging and suggests the sample shows inhibition. The “inhibition” result indicates an obstruction within the sample that prevents the PCR reaction.