Prehybridization Hybridization Solution for Hybridization in Formamide Buffers

5x Denhardt's reagent

100 jg/ml salmon sperm DNA 50% (v/v) formamide

After a thorough mixing, filter the solution through a 0.45-jm disposable cellulose acetate membrane (Schleicher & Schuell Uniflow syringe membrane or equivalent). To decrease background when hybridizing under conditions of reduced stringency (e.g., 20-30% formamide), it is important to use formamide that is as pure as possible.

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