Hi,
We are considering purchasing a Bioanalyser to increase the accuracy of DNA quantification during library preparation for next generation sequencing. However I have heard that some laboratories now use a Tapestation instead.
Given that Tapestations are more expensive, I wondered if anybody had any advice regarding either of these machines and could offer their own recommendations.
We use Bioanalyzer and Qubit or qPCR to quantify our libraries.
With a library prep kit that we use a lot, we have the following calculation to get the molarity:
nM = (Qubit value ng/µL)*106 / ((average size from BioA bp * 607.4) + 157.9)
The Qubit value tends to over estimate because it detects all the DNA fragments (even the ones w/o adapters), so you can dilute a bit more if you are afraid to overload the sequencer.
qPCR remains the more reliable technique as it amplifies only the fragments with adpaters (and adapter dimers too). You can try and compare the two techniques to see if the above calculation is close to the qPCR result (it usually is) and choose the one you prefer (regarding costs, ease of use...)
Good luck!
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