With the Qubit High Sensitivity DNA kit, I was measuring 8 samples (2uL each + 198 of the working solution), and noticed that 4 of the 8 samples had readings of >60ng/uL. I then re-did the Qubit readings with the Broad Range kit and compared the two readings (High Sensitivity vs Broad Range).
2 of the 4 samples that had a reading of >60ng/uL (High Sensitivity) now had readings of 40ng/uL and 46ng/uL (both Broad Range). The other 4 samples (the ones with readings 60ng/uL but, then again with Broad Range, only 2 of those 4 samples were actually ~40ng/uL? Also, when comparing the samples that were already within the range for High Sensitivity, why is there a discrepancy with the Broad Range? Was the Broad Range less accurate for samples with lower DNA concentrations?
Any articles/other posts about the Qubit High Sensitivity and Broad Range limitations would be appreciated. Thank you.
Did you try to do separate replicates of the same samples with each kit to see what the natural variability is? Since you're working with such small volumes of sample (2 µL) it might be something as simple as your pipette delivering slightly different amounts each time. (ie. the difference between 2 µL and 2.5 µL at 40 ng/µL is 20 ng of DNA)
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