PF Tek failure troubleshooting — what went wrong and how to fix it

Green appearing at days 10-12 is almost always Trichoderma, which means contamination entered after sterilization — not during it (Tricho wouldn't survive the pressure cooker). The most common entry points: (1) inoculation technique — the needle didn't cool long enough before injection, or the surface wasn't wiped with alcohol; (2) the polyfill or tyvek in your lid isn't adequate — if moisture wicks into the port, it bypasses your sterile barrier; (3) the jar neck had residue when you lidded — wipe the inside of the jar neck clean before lidding.

Specific tip: after pressure cooking, let jars cool for 24 hours minimum before inoculating. Inoculating warm jars creates condensation inside — moisture on the walls is contamination risk. Also: shake the jars when the mycelium is partial (30-40% colonized) to distribute it evenly — this speeds colonization and reduces the window in which contaminants can establish.

Your glove box sterility may be the issue. Still Air Box technique: let the still air box sit undisturbed for 15-20 minutes before working in it — air movement disturbs particles and then they settle. Also wipe the inside of the SAB with isopropyl before use. If you're moving quickly inside the box, you're stirring air that carries contaminants. Slow down: all movements inside the SAB should be deliberate and unhurried.