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.
摘要:
这项研究旨在开发一种使用脱氧尿苷探针和聚合酶链反应的短DNA检测方法。探针与靶短DNA杂交,然后通过DNA聚合酶延伸。在通过尿嘧啶DNA糖基化酶去除探针之后,将延伸的DNA用于实时PCR。该方法测量了0.01至10nM的17个核苷酸的模型短DNA序列。然后将该方法用于核酸药物fomivirsen的检测,以及21个硫代磷酸酯核苷酸,并量化0.1-100nM的fomivirsen。该方法可用于检测短DNA片段,如功能性核苷酸。