Why do I see multiple peaks in the dissociation curve after my qPCR assay?
There are several reasons for seeing multiple peaks in the real-time dissociation curve. Always verify amplicon production by agarose gel electrophoresis. Larger bands could be due to genomic DNA contamination, which can be verified using a No Reverse Transcription (NRT) control. Smaller bands could be due to the presence of primer dimers. Extra bands could also be due to the presence of un-annotated alternative transcripts or splice variants in your total RNA sample.
Related Questions
- What is a dissociation curve, and why is it particularly important to run a dissociation curve, following qPCR using SYBR Green chemistry?
- Why do I see multiple high-intensity peaks in my qPCR dissociation curve at temperatures less than 70 ºC?
- Why do I see multiple peaks in the dissociation curve after my qPCR assay?
