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Anyone found a 96 well clean up method that will remove 100 nt/bp and smaller from ~250 bp product? Most give 100 bp as the cutoff, but we've gotten too much small material back with Qiaquick 96. Alternative is Ampure, and it can be tricky to handle in 96 well plate. We'd like to clean up, quantitate, THEN pool and size select to get rid of rest of small material before sequencing. Have tried quantitation, pooling, then clean up and size selection, and dataset yields vary too much. Thanks; Hilary
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We've had success with the 96 Well PureLink Pro kit from Invitrogen
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Nothing will outperform AMPure IMHO. You'll probably have to titrate the cut (specifically the binding time and HOW you mix [static vs constant mixing]) in your own hands, but I would start with 1.3-1.4x for that separation range.Comment
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Love this - good data definitely starts from good input, and poor input can only give relatively poor data. I particularly like the mention of Nanodrop/absorbance based methods for quantification. It's such a toss up if you'll get an accurate reading or what amounts to a randomly generated number, and a lot of library/sequencing related issues can be traced back to poor quant.07-01-2026, 11:43 AM
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I’m not a sequencing expert. I’m a purification scientist who uses NGS to evaluate workflows my group develops. With this perspective, we think about the sample first and the NGS workflow second. The sequencer is an exceptionally honest reporter, but it can only report on what you give it, so whether you get clean, interpretable data from an NGS workflow is largely determined before you begin.
Here are nine questions we think about, in roughly the order they matter, before...
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