Table 1.

RADD reaction conditions. RADD is performed in two sequential reactions without aspirating reagents between reactions. The lesion processing mix (left) is placed on prepared tissues and placed in a humidified incubator. The gap-filling mix (right) is added directly to the lesion processing mix and incubated for an additional hour. The reagents are then aspirated, and the cells are washed and incubated with anti-digoxigenin antibody.

Full RADD Lesion Processing Mix	Per 100 µL Reaction Volume	Gap-Filling Mix	Per 100 µL Reaction Volume
UDG (NEB M0280)	2.5 U	Klenow exo- (Thermo Fisher, Waltham, MA, USA, EP0422)	1
FPG (NEB M0240)	4 U	Digoxigenin dUTP (Sigma Aldrich, St. Louis, MO, USA, 11093088910)	0.1
T4 PDG (NEB M0308)	5 U	Thermo Pol Buffer	10 µL
		(NEB B9004)
EndoIV (NEB M0304)	5 U
AAG (NEB M0313)	5 U
NAD+ (100x, NEB B9007)	500 µM
BSA (Sigma Aldrich, St. Louis, MO, USA)	200 µg/mL
Thermo Pol Buffer	10 µL
(NEB B9004)