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#1 Nephrite



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Posted 16 March 2011 - 03:20 AM


Here, in the forum, I read everything about primer dimerization and the possible ways how it could be overpassed - many thanks about this!
Also here, I met the word ''concatamerization'' - a sort of ligation.

I have primers 20b long, and intesive unspecific product of ~ 50 bp in positive and in negative samples, which appears only when both primers are together - i.e. it is missing if I load only R or F. In the latter case, there are very pale traces of ~ 20b.

1. How can I distiguish whether it is a crossdimer, or concatamere and does it matter?

2. What is the reason for concatamerization and could it be overcome by decreasing the polymerase concentration?

So far, I managed to diminish the intensity of this product in my positive samples, but not in NTC. Will this compromise the qPCR, which is my final aim?

Any advice would be of a great help. Thank you.

PS - NTC means no template control, i.e. negative sample with water insted of DNA template, right?

Edited by Nephrit, 16 March 2011 - 03:26 AM.

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