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What does it mean if I see other significant peaks in the melt analysis of my standards?
It is normal to see a slight shoulder on the low-temperature side of the sample melt peak. If the shoulder is very large, either extra Illumina® Primer mix was added or an insufficient amount of standard was added. In this case, the experiment should be repeated with special care taken while making the master mix and when adding standards to the reaction.
If multiple peaks are observed in the melt curves of the standards, then contamination was introduced into the reaction. If it is present in all of the standards, the contamination is in the master mix. The experiment should be repeated with new aliquots of reagents. If it is present in the replicates of only one standard, that standard aliquot has become contaminated and must be discarded.