Table 1.

Troubleshooting Table

Step	Problem	Reason	Solution
19	Efficiency of gDNA extraction is poor.	Not enough tissue was sampled.	Resect more scales from individual animals. Submerge forceps with scales in NaOH lysis solution for 10 second before agitation.
22	Verified prkdc−/−, il2rgα−/− are sick.	Verified prkdc−/−, il2rgα−/− shows signs of infection, such as clamped up fins, lethargy, and/or disorientation.	Animals might be infected from genotyping procedure. Either euthanize animals or move them into an isolation tank and supplement animals with 2X recommended dose of antibiotics for a week. Verify the correct dosing and efficacy of antibiotics is being used.
37	An air bubble was trapped in syringe containing transplant mixture.	Air bubble was introduced during mixing.	Be more cautious and pipette slowly when mixing different components of the transplantation mix.
41	Low engraftment rate	Some cell lines do not engraft well in our prkdc−/−, il2rgα−/− animals	Increase cell number transplanted from 5 × 105 to 2 × 106 cells per animal.
49	Excessive bleeding post IP injection	Needle broke pleural ribs of animal, or was inserted too deep into the intraperitoneal cavity causing injurty to internal organs.	Make sure to insert the needle between ribs and stop inserting needle once the bevel has fully entered the cavity.
52	High mortality post imaging of engrafted cells	Imaging time was too long or tricaine dosing was incorrect.	Limit procedure time to a maximum of 5 min per animal when imaging. Use 0.1 mg/ml tricaine to anesthetize animal.
53	Drugs are expelled from the gills during oral gavage	The oral gavage catheter was not inserted deep enough into the animal or drugs were delivered to fast.	Make sure the rubber tip of feeding catheter is inserted completely into the animal, with the animal vertically oriented, with mouth and syringe aligned in a straight line. Inject slowly.