Vincent U, Patra G, Therasse J, Gareil P
Service des Biotechnologies, CEB, Vert-le-Petit, France.
Electrophoresis. 1996 Mar;17(3):512-7. doi: 10.1002/elps.1150170316.
Quantitation of DNA fragments amplified by polymerase chain reaction (PCR) is needed for the determination of target DNA in molecular biology. Capillary electrophoresis in entangled polymer solution coupled to laser-induced fluorescence detection was assessed as an alternative technique to conventional slab gel methods to monitor competitive PCR, which consists of amplifying an internal standard fragment under the same conditions as the target fragment. The fluorescence signal was generated either through end-labeling of the fragments using 5'-fluorescein-labeled primers or through intercalation of ethidium bromide along the double strand. It is shown that the more accurate and reliable results were obtained using this latter pathway.
在分子生物学中，为了测定目标DNA，需要对通过聚合酶链反应（PCR）扩增的DNA片段进行定量分析。评估了在缠结聚合物溶液中进行的毛细管电泳与激光诱导荧光检测相结合的方法，将其作为传统平板凝胶方法的替代技术，用于监测竞争性PCR，竞争性PCR是指在与目标片段相同的条件下扩增内标片段。荧光信号可以通过使用5'-荧光素标记的引物对片段进行末端标记产生，也可以通过溴化乙锭沿双链插入产生。结果表明，使用后一种方法能获得更准确可靠的结果。
