AP站点
化学
检出限
限制
DNA
基底切除修复术
末端脱氧核苷酸转移酶
分子生物学
核酸内切酶
组合化学
计算生物学
DNA修复
生物化学
色谱法
细胞凋亡
标记法
工程类
生物
机械工程
作者
Zhe Hu,Weicong Ye,Zhen Zhang,Tianci Xie,Wenqian Yuan,Tongbo Wu
标识
DOI:10.1016/j.aca.2022.340220
摘要
The apurinic/apyrimidinic (AP) site is one of the most common DNA lesions and a critical intermediate during the base excision repair pathway. Therefore, AP sites are essential in clinical diagnosis, treatment and detection. However, the existing detection methods are complicated in design and synthesis and have high instrument requirements, limiting their wide application. Therefore, there is an urgent need for a sensitive and straightforward detection method without time-consuming and heterogeneous reactions. Herein, we developed two compatible detection methods for AP sites in long and short dsDNA. For long and short dsDNA, the background signal was successfully suppressed by the affinity difference of Terminal deoxynucleotidyl transferase (TdT) and 3' -end blocking, respectively, thus achieving high detectability and specificity. The detection limit was 13 pM in 20 μL, meaning that the LOD was 0.26 fmol for AP site amount and 0.05% for AP site abundance. The method has been successfully applied to detect AP sites in various biological samples quickly. Therefore, it has broad clinical application prospects, catering for the need for a point of care.
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