Nishikawa T, Kambara H
Central Research Laboratory, Hitachi Ltd., Tokyo, Japan.
Electrophoresis. 1994 Feb;15(2):215-20. doi: 10.1002/elps.1150150136.
Long DNA fragments were separated by capillary gel electrophoresis with laser-induced fluorescence detection, and two approaches to obtaining high-resolution separation of DNA fragments were examined. One method achieved high resolution at the expense of detection time, while the other achieved both high resolution and fast detection. The former approach used a 300 cm long capillary with a gel concentration of 4% T and an electric field strength of 70 V/cm. The resolution limit in this electrophoresis was 800 bases with a resolution value of 0.5 for adjacent peaks. Under these conditions, the measurement time was 44 h. The latter approach used a 200 cm long capillary with a gel concentration of 3% T and an electric field strength of 170 V/cm. The resolution limit in this electrophoresis was 680 bases with a resolution value of 0.5 for adjacent peaks, and the measurement time was only 10 h. With both approaches, fragments with less than 600 bases were efficiently separated. The resolution values for adjacent peaks with less than 500 bases are greater than 1.0, and those for peaks with less than 100 bases are greater than 2.7. These approaches thus improve the accuracy of the base sequence determination.
长DNA片段通过毛细管凝胶电泳结合激光诱导荧光检测进行分离，并研究了两种实现DNA片段高分辨率分离的方法。一种方法以牺牲检测时间为代价实现了高分辨率，而另一种方法则同时实现了高分辨率和快速检测。前一种方法使用了一根300厘米长的毛细管，凝胶浓度为4%T，电场强度为70V/cm。这种电泳的分辨率极限为800个碱基，相邻峰的分辨率值为0.5。在这些条件下，测量时间为44小时。后一种方法使用了一根200厘米长的毛细管，凝胶浓度为3%T，电场强度为170V/cm。这种电泳的分辨率极限为680个碱基，相邻峰的分辨率值为0.5，测量时间仅为10小时。使用这两种方法，小于600个碱基的片段都能得到有效分离。小于500个碱基的相邻峰的分辨率值大于1.0，小于100个碱基的峰的分辨率值大于2.7。因此，这些方法提高了碱基序列测定的准确性。
