%0 Journal Article
%T Real-time PCR method for detection of short DNA using a deoxyuridine probe and application for detection of fomivirsen.
%A Harikai N
%A Tanaka Y
%A Miyashita S
%A Zaima K
%A Shinomiya K
%J Biotechniques
%V 73
%N 6
%D 12 2022
%M 36350046
%F 2.746
%R 10.2144/btn-2022-0068
%X This study sought to develop a short DNA detection method using a deoxyuridine probe and polymerase chain reaction. The probe was hybridized to the target short DNA, which was then extended by DNA polymerase. The extended DNA was used for real-time PCR after the probe was removed by uracil DNA glycosylase. This method measured from 0.01 to 10 nM of a model short DNA sequence of 17 nucleotides. The method was then used to detect the nucleic acid medicine fomivirsen, as well as 21 phosphorothioate nucleotides, and to quantify 0.1-100 nM of fomivirsen. This method may be useful for detecting short DNA fragments, such as functional nucleotides.