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We are using the Chemagen system (http://www.chemagen.com/fileadmin/do...n_brochure.pdf) to extract total nucleic acid from blood and stool samples. The process yields a mixture of purified DNA/RNA, but doesn't suggest how to separate the two. Has anyone done this type of separation before? I know Qiagen uses columns in their AllPrep system, but they can't be purchased separately. Most RNAse or DNase protocols I see are designed to remove small amounts of contaminating RNA/DNA, I don't know how they'd do with total nucleic acid?
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If you need both RNA & DNA from the same prep, then split your prep in half. Treat one half with DNase, then run it over a column (qiagen/zymo whatever) to clean up. Base hydrolyze the other half, then EtOH precipitate. -
Thank you very much, that's likely how I'll proceed. I was going to use RNase A, but base hydrolysis is likely more cost effective, especially for a lot of samples.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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