Reagent	Quantity (for 50 ml)	Final concentration
Deionized formamide	25 mL deionized formamide	50%
SSC	12.5 mL of 20× SSC	5×
Heparin	2.5 mg heparin	50 μg/mL
Tween-20	50 μl 100% Tween-20	0.1%
Adjust the pH to 5.0 using HCl and bring the final volume to 50 ml with sterile dH20. Hyb buffer is stored at −20° C.
For Hyb-DNA-SDS, combine 14.85 ml Hyb solution and 150 μl of 10 mg/ml sonicated salmon sperm DNA (Invitrogen Cat. No. 15632-011). Store at −20° C. Prior to use, warm the solution and add SDS to a final concentration of 0.3%. Use this solution in the blocking step prior to hybridization and during the actual hybridization.