I have been encountering false positive results while processing the samples for influenza typing and subsequent subtyping. I was wondering if real time rt pcr (without calibration) could be the possible reason behind it because i have tried almost everything as a part of troubleshooting.. need suggestions.
Yes, sure. Cross contamination can be an important source of false positive results for real-time PCR.
Dear Hyder, please, try the following trouble shooting:
1. Do a general cleaning and decontamination of your laboratory and the facilities using 5% hypochlorite and re-run the samples.
2. Adhere to PPEs, hand gloves, nose mask, hair cover, etc, to prevent all contaminants.
3. Run your controls, check your buffers, etc, to be sure they are not contaminated.
Cheers.
Good, I am happy that you followed and every thing is now working.
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