{Reference Type}: Journal Article
{Title}: Real-time PCR method for detection of short DNA using a deoxyuridine probe and application for detection of fomivirsen.
{Author}: Harikai N;Tanaka Y;Miyashita S;Zaima K;Shinomiya K;
{Journal}: Biotechniques
{Volume}: 73
{Issue}: 6
{Year}: 12 2022
{Factor}: 2.746
{DOI}: 10.2144/btn-2022-0068
{Abstract}: 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.