School of Pharmaceutical Science and Technology, Tianjin University, Tianjin 300072, China.
Tianjin University and Health-Biotech United Group Joint Laboratory of Innovative Drug Development and Translational Medicine, Tianjin University, Tianjin 300072, China.
Biomed Res Int. 2023 Jan 23;2023:8296847. doi: 10.1155/2023/8296847. eCollection 2023.
DNA detection, which includes the measurement of variants in sequences or the presence of certain genes, is widely used in research and clinical diagnosis. Both require DNA-dependent DNA polymerase-catalyzed strand extension. Currently, these techniques rely heavily on the instruments used to visualize the results. This study introduced a simple and direct colorimetric method to measure polymerase-directed elongation. First, pyrophosphate (PPi), a by-product of strand extension, is converted into phosphate (Pi). Phosphate levels were measured using either Mo-Sb or BIOMOL Green reagent. This study showed that this colorimetry can distinguish single-base variants and detect PCR products in preset stringent conditions, implicating the potential value of this strategy to detect DNA.
DNA 检测，包括序列变异的测量或特定基因的存在，广泛应用于研究和临床诊断。这两者都需要 DNA 依赖性 DNA 聚合酶催化的链延伸。目前，这些技术严重依赖于用于可视化结果的仪器。本研究引入了一种简单直接的比色法来测量聚合酶指导的延伸。首先，链延伸的副产物焦磷酸（PPi）转化为磷酸（Pi）。使用钼-锑或 BIOMOL Green 试剂测量磷酸水平。本研究表明，这种比色法可以区分单碱基变异，并在预设的严格条件下检测 PCR 产物，这表明该策略在检测 DNA 方面具有潜在价值。
