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Does anyone have feedback on the Kapa HyperPlus enzymatic DNA fragmentation vs Covaris? I have access to a Covaris but the enzymatic method would be much more convenient... assuming it works well!
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Almost all suppliers in their technotes claim that their enzymatic DNA fragmentation gives similar or better results than Covaris or their competitors. My comments:
1- Shearing is not sensitive to buffer or DNA quality to some extent, but all the enzymatic methods require or prefer a particular buffer for best results.
2- Buffer requirement is really annoying for core centres dealing with DNA from different clients
3- The enzymatic methods are time or quantity sensitive requiring special attention
4- These methods are acceptable for labs that do not have access to Covaris or desire fast high throughput fragmentation -
I agree with nucacidhunter. Covaris shearing will produce much more consistent size distributions across all your samples. When I tried to use KAPA fragmentation enzyme, size distributions were highly sensitive to the input DNA concentrations. Samples with small DNA concentrations (<100ng) required much shorter time for fragmentation than those with ~500ng. And samples with DNA concentration >750ng always produced libraries with a large portion of long fragments. Therefore, be prepared to spend some time and reagents for calibration before you actually start doing your libraries with enzymatic fragmentation.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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