genomic DNA quantification

[Rocketknight]

Picogreen or Qubit should be sufficiently accurate. The Nanodrop and Bioanalyzer will often give more varying results. The Nanodrop tends to overestimate the DNA concentration due to contamination with ssDNA/nucleotides/RNA/etc. The Bioanalyzer depends on accurately pipetting very small volumes, so it's always a bit tricky.

In my experience, those three methods frequently disagree with each other, but measurements with Qubit on the same sample at different times have given fairly good reproduction.

[Roxanne Kelly]

My lab is a biobank that sends samples out to external labs for next gen sequencing. However, we find it nearly impossible to get matching results from our internal quantification methods and the lab that we are sending to.

My lab uses picogreen quantification to quantify the input genomic DNA, we also have a Nanodrop and an Agilent Bioanalyzer. We have found that our 3 pieces of equipment often disagree with each other and of course our measurements disagree with external lab's measurements. Most labs that perform sequencing double-check the DNA concentration prior to running and require a certain range in order to "pass" a sample in their QC. And of course everyone thinks that their measurements are right and everyone else's are wrong. Very frustrating.

Does anyone know of a better, more accurate measurement for DNA than the 3 listed above? I feel like since NextGen is so picky about this, there needs to be a better method?

Thanks,
Roxanne.