So I did a real time PCR lately to investigate the CT/copy number value of cDNA after doing a reverse-transcriptase PCR of pLVX-myc mRNA extracted from about 250 ng of MSC-derived exosomes. The results were not exactly according to my expectations and I wanted to find out if there's anything in the exosome sample that is inhibiting the real-time PCR.
My boss has suggested a spiking when doing the real-time PCR. I do not understand the principle and concept of doing this. Can someone kindly explain to me clearly what is it and why is it done for certain PCR reactions?
If I am not mistaken, "spiking" here is equivalent to adding a "spike" control.