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Why is my Ion AmpliSeq library concentration high? Product FAQ


Here are possible causes and solutions:
- The sample DNA or RNA may have been misquantified. We recommend requantifying the DNA or RNA. See the user guide for your library preparation method.
- More than 100 ng of sample DNA or RNA may have been used. We recommend decreasing the number of amplification cycles by adding less DNA or RNA.
- If you are using the Agilent 2100 Bioanalyzer instrument, the markers may have been misassigned. We recommend ensuring that the markers are correctly assigned.
- The library may have been contaminated by high molecular weight DNA. We recommend one of the following:
a) Removing less supernatant in the first-round (0.5X) purification and to ensure that the bead pellet is not disturbed.
b) Increasing the AMPur XP Reagent volume from 25 µL (0.5X) to 35 µL (0.7X) in the first-round purification.
- Inserts may have concatemerized during the ligation steps. We recommend reducing nucleic acid input, requantifying samples with a Qubit Fluorometer, or reducing the target amplification cycle number.

Answer Id:: E18935

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