Mitnik Luba, Carey Loucinda, Burger Robert, Desmarais Samantha, Koutny Lance, Wernet Oliver, Matsudaira Paul, Ehrlich Daniel
Whitehead Institute for Biomedical Research, Cambridge, MA 02142, USA.
Electrophoresis. 2002 Mar;23(5):719-26. doi: 10.1002/1522-2683(200203)23:5<719::AID-ELPS719>3.0.CO;2-K.
We report the development of a robust and effective method for multiplexed short tandem repeat (STR) analysis within a chip-based microdevice. The method uses a laser-induced fluorescence detection system and simultaneously detects three- and four-color multiplexed polymerase chain reaction (PCR) samples. Analyses of the eight combined DNA index system (CODIS) STR loci were performed in 20 min with single-base-pair resolution ranging from 0.75 to 1. A simultaneous analysis of fifteen loci-ladders and a gender marker Amelogenin based on the PowerPlexTM 16 System was achieved in less than 35 min. The system is capable of repetitive operation and may be extended to high-throughput multilane devices that could be readily interfaced to an automated sample loading system.
我们报告了一种在基于芯片的微器件内进行多重短串联重复序列(STR)分析的强大且有效的方法的开发。该方法使用激光诱导荧光检测系统,同时检测三色和四色多重聚合酶链反应(PCR)样品。在20分钟内对八个联合DNA索引系统(CODIS)STR位点进行了分析,单碱基对分辨率范围为0.75至1。基于PowerPlexTM 16系统,在不到35分钟的时间内实现了对十五个基因座阶梯和性别标记物牙釉蛋白的同时分析。该系统能够重复操作,并且可以扩展到可轻松连接至自动样品加载系统的高通量多通道器件。