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.

Biosafety take: none of this is a problem at BSL-1 with non-pathogenic chassis. The failure mode in community labs is almost never the organism, it is the chemical waste.

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.

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?

It is in there — line 14, the $22 board. The expensive part historically was the Peltier driver and that is what got cheap.