Blokhin Russian Cancer Research Center, Russian Academy of Medical Sciences, Kashirskoe Shosse 24, 115478 Moscow, Russia.
Anal Biochem. 2011 Jan 15;408(2):304-8. doi: 10.1016/j.ab.2010.09.004. Epub 2010 Sep 7.
The technique of isotachophoresis is intended for separation of molecules having different electrophoretic mobilities in a nonhomogeneous electric field. Since the mobility of nucleic acids in water solutions is uniform and does not depend on their size (because of a uniform distribution of negatively charged phosphate groups along the molecule), isotachophoresis will concentrate rather than separate them in the mobile borderline zone between the rapid (Cl(-)) and the slow (β-alanine(-)) anions. This idea served as the basis for elaboration of a novel method for isolation of nucleic acids from diluted solutions. Advantages of the method include quantitative yield (regardless of molecule size), high degree of concentration, and the ability to visually monitor the process. The method may find applications in nucleic acid isolation from highly degraded forensic and clinical samples, from bodily fluids in particular, and thereby promote development of this important direction of diagnostics.
等速电泳技术旨在用于分离在非均相电场中具有不同电泳迁移率的分子。由于核酸在水溶液中的迁移率是均匀的,并且不依赖于它们的大小(因为带负电荷的磷酸基团在分子中均匀分布),因此等速电泳将在快速(Cl(-))和慢速(β-丙氨酸(-))阴离子之间的移动边界区域集中而不是分离它们。这个想法成为从稀释溶液中分离核酸的新方法的基础。该方法的优点包括定量产率(与分子大小无关)、高浓缩度以及能够可视化监测过程。该方法可应用于从高度降解的法医和临床样本中分离核酸,特别是从体液中分离核酸,从而促进该重要诊断方向的发展。