N-Allyl-4-aminobenzenesulfonam

Revised amino-allyl labeling protocol

for an additional 1 hour.     B. Hydrolysis     1. Degrade RNA by addition of 15 m L of 0.1 N NaOH. Incubate at 70C for 10 minutes   2. Neutralize by addition of 15 m L 0.1 N HCl.     To continue with the amino-allyl dye coupling procedure

Amino-allyl Reverse Transcription

ul 1N NaOH 10ul .5M EDTA Incubate: 15 min. at 65° C. Neutralize: 25ul 1M Tris pH 7.4 or 1M HEPES pH 7.5 III. Cleanup To continue with the amino-allyl dye coupling procedure all Tris must be removed from the reaction to prevent

Protocol for Reverse Transcription and Amino-allyl Coupling of RNA

by addition of 15 mL 0.1 N HCl.    To continue with the amino-allyl dye coupling procedure, all Tris must be removed from the reaction to prevent the monofunctional NHS-ester Cy-dyes from coupling to free amine groups in solution.   3.  Add 450