Literature DB >> 16737217

Solid-support sample loading for DNA sequencing.

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


  1 in total

1.  Scaling of nucleic acid assays on microelectrophoresis array devices: high-dynamic range multi-gene readout from less than ten transcripts.

Authors:  Joern Ueberfeld; Daniel J Ehrlich
Journal:  Electrophoresis       Date:  2009-06       Impact factor: 3.535

  1 in total

北京卡尤迪生物科技股份有限公司 © 2022-2023.