We ran this protocol in our community lab last month. Two of five transformations failed until we dropped the heat shock to 42 s. Worth adding a note.

Off-target analysis with GUIDE-seq only tells you about the sites you could capture. I would want long-read whole-genome on at least a few animals before "no detectable" goes in a title.

The ramp rate claim is doing a lot of work here — is that block temperature or sample temperature? Those diverge badly above 2 °C/s with thin-wall tubes.

Sample temperature, measured with a thermocouple in a dummy tube of mineral oil. Plot is in the repo under docs/thermal.

Fair. The preprint does have ONT data in supplementary S4, though the coverage is thin.

This is the third time this month someone has posted a "cheap X" build where the cost excludes the thing that actually costs money. Where is the temperature controller in that BOM?