Problem	Potential causes/solutions
Tranforming pDawn-Ag43 into host strain - no colonies	Low plasmid concentration - check plasmid concentration on spectrometer. A typical miniprep of pDawn-Ag43 should yield at least 100 ng/μL; use up to 10-100 ng for transformation
Check/remake competent cells: competent cells should have transformation efficiency at least 10^6 cfu/µg verified using a standard plasmid such as pUC19 - if not, remake competent cells
Wrong (level of) antibiotic on LB agar plate - make sure to use 50 μg/mL spectinomycin  for selection
Projector illumination turns off / inconsistent overnight	Disable problematic software such as: automatic overnight software/OS updating, night-time blue-light filter
Projector may be overheating - set incubator to lower temperature while projector is turned on (e.g. 30 °C instead of 37 °C) - note projector as heat source can overheat incubator beyond set point
Remove unnecessary sources of humidity from incubator, as these may affect projector electronics
No/low levels of biofilm formed after overnight illumination, no  planktonic cells growth either (i.e. liquid is clear)	Wrong (level of) antibiotic  - make sure to use 50 μg/mL spectinomycin
Check everything is added to M63 recipe properly
No/low levels of biofilm formed after overnight illumination, only planktonic cells (i.e. liquid is cloudy)	Check light level, projector should be illuminating blue light at 50 μW/cm^2 at 460 nm wavelength
Try letting bacteria grow for shorter/longer time after 1:1000 LB subdilution step prior to adding to M63
Restreak bacteria on LB plate, start from fresh colony to generate overnight stationary phase culture
Ensure projector is working consistently overnight - see point above
Fuzzy biofilm patterns, high levels of background noise	Reduce stray light from optical illumination system, cover reflective surfaces on interior of incubator
Consider using photomask-based (as opposed to projector-based) structured illumination
Check projector is properly focused at the bottom surface of the biofilm culture chamber