| Literature DB >> 16737217 |
Jörn Ueberfeld1, Sameh A El-Difrawy, Korisha Ramdhanie, Daniel J Ehrlich.
Abstract
We present a new method for simplified low-quantity DNA loading onto microelectrophoresis devices. The method is based on combined solid-phase extraction, purification, and transport of DNA reversibly bound on paramagnetic microspheres. DNA is adsorbed onto the microspheres, captured with a magnetized permalloy wire, and then directly injected as a highly focused sample plug into the separation channel. This method circumvents both the minimum volume requirement of pipettors (since only solid beads are transported) and the timing complications of double-T microfluidic injection. Injections from Sanger samples of <100 pg total suspended weight match the signal strength of our previous conventional injections at >10-times the starting DNA sample. Sequencing traces show a resolution that matches or exceeds double-T injections. A kinetic model reproduces the time-dependence of the injection signals and proves that total nonidealities in the method produce injection-broadened plugs of approximately 1-s duration. The method should be broadly extendable to DNA and protein separations in both microdevice and capillary electrophoresis.Mesh:
Substances:
Year: 2006 PMID: 16737217 DOI: 10.1021/ac052201x
Source DB: PubMed Journal: Anal Chem ISSN: 0003-2700 Impact factor: 6.986