We’re using SYBR green qPCR master mix from Fermentas to check the expression level of our transfected cell lines. I’m getting a signal in my template free negative control (nuclease free water instead of template) after 23th cycle. I also confirmed it on gel. I have changed supply of nuclease free water and purchased new primers and still get a signal. I couldn’t find what is wrong.
Any help would be appreciated.
real time pcr amplification of non-template controls?