Distribution of single DNA molecule electrophoretic mobilities in semidilute and dilute hydroxyethylcellulose solutions

✍ Scribed by Yoshinori Yamaguchi; Todor I. Todorov; Michael D. Morris; Ronald G. Larson

• Publisher: John Wiley and Sons
• Year: 2004
• Tongue: English
• Weight: 122 KB
• Volume: 25
• Category: Article
• ISSN: 0173-0835
• DOI: 10.1002/elps.200305783

✦ Synopsis

Abstract

The distribution of center of mass electrophoretic mobility mobilities and normalized migration time of up to 1080 λ DNA molecules per experiment were measured in both semidilute hydroxyethylcellulose HEC/0.5× Tris‐borate‐EDTA (TBE) solutions and dilute HEC/0.5×TBE solution by high‐speed video microscopy. Measurements were made microscopically over a short migration distance in homogeneous DNA HEC/0.5×TBE solution and after electrophoretic migration of a plug of DNA through 7 cm. Video at 120 frames/s (semidilute HEC solution) and 236 frames/s (dilute HEC solution) allowed visualization with adequate resolution for single molecule mobility measurements. The electrophoretic migration times and band shapes predicted from the measurements corresponded well with those measured by conventional capillary electrophoresis (CE) in both semidilute and dilute HEC. In semidilute solution, the band width predicted by a square root of time scaling is in good agreement with the results of conventional CE. However, in dilute solution the precision of the measurements was not good enough to allow scaled estimates of band widths.