Hello
I Performed real time pcr for 2 blood samples (cDNA) and got the attached results.
1. Why i can't see the Plateau line?
2. Is this result acceptable?
3. is the melt curve correct? and indicates that the amplification was only for my specific amplicon? not primer dimer..... etc?
I didn't run a control or a standard curve, I just wanted to test the amplification of my cDNA
thanks
1. because your PCR is terminated befor amplification has reached the plateau
2. no
3. no. it indicates, that you have traces amplicons of low melting
Yeah, your qPCR did not work. I can't tell you why from this. Definitely run it again with a standard curve. That will tell you more if it is an issue with your samples or your master mix.
By looking your amplification curve I can tell you must check first standard curve so that it will tell you about primer efficiency as well as CT value for quantification. the second thing you can do is dilute cDNA concentration in the reaction mixture and run the triplicate sample. with No template control (NTC). NTC should not have any amplification, so just check it. you can read available literature on the internet, I am sharing you a link, just check it.
http://www.nature.com/nprot/journal/v3/n6/full/nprot.2008.73.html
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