High throughput purification of amino-modified oligonucleotides and its application to novel detection system of gene expression.
We report here a new amino-modifier reagent, which enable high-throughput purification of amino-modified oligonucleotides. Either monomethoxytrityl (MMT) or trifluoroacetyl (TFA) group has been used as the protecting group for the primary amine when amino-terminal oligonucleotides are prepared. Generally, the removal of MMT requires the stringent acidic treatment after the oligonucleotide synthesis. We chemically synthesized a novel amino-modifier with the MMT protection. It was found that the new amino-modifier released MMT group under mild acidic condition, and then rapid purification of diverse amino-modified oligonucleotides could be achieved with cartridge column of reverse phase. Furthermore, we tried to construct the new detection system to study gene expression using the amino-modified oligonucleotides. The new amino-modifier will be useful for molecular biology by facilitating the construction of oligonucleotide library and improving the chemical reactivity.
Kojima
Naoshi
N
Research Institute of Genome-based Biofactory, National Institute of Advanced Industrial Science & Technology (AIST Hokkaido), 2-17-2-1 Tsukisamu-Higashi, Toyohira-ku, Sapporo 062-8517, Japan.
Sugino
Maiko
M
Mikami
Akiko
A
Nonaka
Ken
K
Fujinawa
Yumi
Y
Ueda
Yoji
Y
Sato
Kousuke
K
Ohtsuka
Eiko
E
Matsubara
Kenichi
K
Komatsu
Yasuo
Y
D000588:Amines / Q000737:chemistry; D020869:Gene Expression Profiling; D007202:Indicators and Reagents; D020411:Oligonucleotide Array Sequence Analysis; D015345:Oligonucleotide Probes / Q000138:chemical synthesis / Q000737:chemistry* / Q000302:isolation & purification*
D000588:Amines; D007202:Indicators and Reagents; D015345:Oligonucleotide Probes