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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!
|dimer , pcr primer dimer , primer|
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