Hello, recently I performed real time PCR and after analysis I get a 700000 fold bigger expression than in the control cDNA. Everything was fine with my probes (same concentration etc.) and with my analysis. Is this possible?
Yes it is possible, but you need to tell more about your treatment and conditions
So, in my tested cells were 700000 particles of one type mRNA and 1 particle mRNA in control cells? I had co-culture cancer cells (cc + some cells) as test and the same cc in monoculture as control. After 3 days incubation (standard conditions) I get RNA from cc. Cells were seperated by small size pore membrane.
Do you use relative or absolute quantification? In relative quantification such things come also when the PCR efficiency is not taken into account.
This is relative quantification, I'm use delta delta ct method and realize this is very nice result for me. Tahnk you for answers.
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