Preparation of DNA and RNA Fragments Containing Guanine N2 -Thioalkyl Tethers

Abstract

This article describes procedures for preparation of deoxyguanosine and guanosine derivatives in which the guanine N² contains a thiopropyl tether, protected as a tert-butyl disulfide. After incorporation into a DNA or RNA fragment, this tether allows site-specific cross-linking to a thiol of a protein or another nucleic acid. © 2023 The Authors. Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: Preparation of diisopropyl-1-(tert-butylthio)-1,2-hydrazinedicarboxylate (4) Basic Protocol 2: Preparation of the 2'-deoxyguanosine N² -propyl-tert-butyl disulfide phosphoramidite (12) Basic Protocol 3: Preparation of the guanosine N² -propyl-tert-butyl disulfide phosphoramidite (20) Basic Protocol 4: Preparation of DNA fragments containing N² -propyl-tert-butyl disulfide guanine Alternate Protocol: Preparation of RNA fragments containing N² -propyl-tert-butyl disulfide guanine Basic Protocol 5: Conversion of N² -propyl-tert-butyl disulfide to the free thiol, disulfide 5-thio-2-nitrobenzoic acid disulfide, or ethylamine disulfide.

Keywords: disulfide cross-links; oligonucleotides.