Li X C, Thilly W G
Division of Toxicology, Massachusetts Institute of Technology, Cambridge 02139, USA.
Electrophoresis. 1996 Dec;17(12):1884-9. doi: 10.1002/elps.1150171213.
A serious limitation of capillary electrophoresis in separating mutant from wild-type DNA sequences is the amount of DNA which may be loaded on narrow-bore columns. Because we sought to examine mutant sequences derived from more than 10(9) human cells, we were obliged to overcome this limitation. Thus we examined the feasibility of using wide-bore capillaries of 250, 320 and 540 microns in constant denaturant capillary electrophoresis (CDCE) using conditions which successfully limited the effect of Joule heating. Excellent separations were observed using the wide-bore capillaries comparable to those obtained using a 75 microns capillary. DNA loading capacities were increased almost 100-fold as the bore of capillary increased from 75 to 540 microns.
毛细管电泳在分离突变型和野生型DNA序列时的一个严重局限在于可加样到窄内径柱上的DNA量。由于我们试图检测源自超过10⁹个人类细胞的突变序列，我们不得不克服这一局限。因此，我们在恒定变性毛细管电泳（CDCE）中，使用成功限制焦耳热效应的条件，研究了使用250、320和540微米宽内径毛细管的可行性。使用宽内径毛细管观察到了与使用75微米毛细管获得的分离效果相当的出色分离。随着毛细管内径从75微米增加到540微米，DNA加样量增加了近100倍。
