Open AccessPreparation of DNA Containing 5‐Hydroxymethyl‐2′‐Deoxycytidine Modification Through Phosphoramidites with TBDMS as 5‐Hydroxymethyl Protecting Group
Author(s) -
Dai Qing,
He Chuan
Publication year - 2011
Publication title -
current protocols in nucleic acid chemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.306
H-Index - 17
eISSN - 1934-9289
pISSN - 1934-9270
DOI - 10.1002/0471142700.nc0447s47
Subject(s) - phosphoramidite , hydroxymethyl , protecting group , oligonucleotide , nucleic acid , chemistry , dna , combinatorial chemistry , stereochemistry , biochemistry , organic chemistry , alkyl
This unit describes procedures for preparation of two phosphoramidite building blocks III and IV , both containing a TBDMS as 5‐CH 2 OH‐protecting group. Phosphoramidites III and IV allow efficient incorporation of 5‐hmC into DNA and a “one‐step” deprotection procedure to cleanly remove all the protecting groups. A “two‐step” deprotection strategy is compatible with ultramild DNA synthesis, which enables the synthesis of 5hmC‐containing DNA with additional modifications. Methods are also presented for their incorporation into oligonucleotides by solid‐phase synthesis, subsequent deprotection, and HPLC analysis. Curr. Protoc. Nucleic Acid Chem . 47:4.47.1‐4.47.18. © 2011 by John Wiley & Sons, Inc.