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-   -   Primer dimer or not?! (http://www.molecularstation.com/forum/biology-forum/85057-primer-dimer-not.html)

Lorna 04-03-2012 01:17 PM

Primer dimer or not?!
 
Ok so I firstly tested out my primer pair I had ordered on a few samples inititally, lovely clean amp curves, clean melt curves with one peak etc.. I then purified the product to make a standard as I work form the standard curve qPCR method. The standards are amplifying nicely but there is a second peak in the melt curve before the main peak. I have no signal AT ALL in my non template controls so I am confused as to if these are primer dimers when they initially gave no evidence of having any? I also changed all my reagents and its still occurring!Any help or advice would be appreciated!
L:headbonk:


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