I am currently running absolute quantification PCR on DNA samples and I use the 3-step cycling steps recommended by the manufacturer (Bioline). Their thermocyclic conditions does not suit me, including the melt curve analysis. I am still optimising for 2 weeks now (sighs!!)
Can anyone share his or her protocol with me.
Thank you
Navonil Gogoi
According to me the problem here is the High Rox Concentration. The instruments like the Biorad CFX96 doesn't require Rox normalization. You should go for the SensiFast No-Rox kit. You can go through this link
https://international.neb.com/faqs/2015/05/18/which-rox-should-i-use-for-my-qpcr-machine
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