Table 2.

Troubleshooting and practical advice.

Step	Potential problem	Cause	Solution
Common bile duct injection	Uncomplete injection of the pancreas	Duct perforation	Continue digestion according to the protocol (you should obtain a good yield)
Pancreas digestion	Partial digestion of caerulein-treated pancreas	Fibrosis may make the tissue more resistant	Incubate an additional 5-to-10 min with Col-Ca2+ buffer at 37°C
	Cell mortality	High collagenase activity/Prolonged incubation	Check collagenase batch/reduce time of digestion to 5 min during the second incubation with Col-Ca2+ buffer
Flow Cytometry	Low number of sorted cells	False gating parameters/Naturally low number of cells	Ensure that the gate covers all YFP-positive events/sort two pancreas digested separately in one tube
	Stuffy nozzle	High number of small debris/DNA rings formation	Use fresh buffers
	YFP-positive cells shift toward more PI staining	Cell damage during sorting	Reduce pressure and drop frequency
RNA extraction and quality	Some samples have remarkably high RNA contamination	Filter may flip over	Avoid touching the filter with tips
	Undetectable RNA bands	RNA is not well-mixed with the gel	Vortex the microfluid chip for 1 min