Hybridization Buffer for ISH

Recipe

Reagent	Amount for One Reaction
Deionized formamide	16 µL
SSPE (20×, pH 7.4)	3.2 µL
tRNA (Type X; 20 µg/µL; Invitrogen/Life Technologies 15401-011)	3.2 µL
Bovine serum albumin (BSA; 20 µg/µL)	1.6 µL
Polyadenylic acid potassium salt (20 µg/µL)	1.6 µL
DEPC-treated H₂O	to 30 µL
Prepare fresh at room temperature. Use solutions prepared with certified nuclease-free H₂O or DEPC-treated H₂O; ultrapure nuclease-free 20× SSPE can be purchased from several vendors. Use Type X tRNA; Types V and X-SA tRNA can contribute to high background.