Hello everyone!
I am using TaqMan probe-based Real-time PCR for the detection of typhoidal salmonella. Since I am not concerned about the expression level of gene, I just want to detect if there is any typhoidal DNA present in my sample or not. Do I still need to add internal control in my reactions?
If yes, then should I add the primer and probes for the housekeeping gene in every reaction since I am doing multiplex PCR or can I add it in a few reaction tubes in every run separately?
Yes, it is recommended to include an internal control in your TaqMan probe-based Real-time PCR assay to monitor the performance of the assay, detect potential issues, and ensure the reliability of the results. You can either design the internal control to work in a multiplex PCR with your target DNA or run separate reactions for the target DNA and the internal control.
good luck,
Djihad Chenna
If you run the reaction in a separate tube, its no longer 'internal'.
If its from human samples, then use the RNaseP assay in a ROX or Cy5 channel (no cross reactivity with FAM, presumably your salmonella assay).
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