Micropreparative capillary electrophoresis of DNA by direct transfer onto a membrane

Electrophoresis
S MagnúsdóttirJean-Louis Viovy

Abstract

We have developed a new technique for the collection of DNA fragments separated by capillary electrophoresis, by direct transfer from the capillary outlet to a positively charged membrane. Transfer and post-run detection of two different nonradioactively labeled DNA standards, ranging in size from 150 bp to 2 kbp and 120 bp to 23 kbp are presented, and discussed. Capillary electrophoresis with direct blotting presents several advantages over the blotting from gels: the separation is faster and requires less manual steps, the resolution is higher, and each DNA fragment is collected into a very concentrated spot on the membrane due to the small surface of the capillary outlet and to a design of the collection device inducing a refocusing of field lines across the hybridization membrane. Therefore, very small amounts of DNA (in the pg range) can be detected. This fraction collection makes further analysis of the sample possible, e.g. by hybridization, thus suppressing one of the major present limitations of the capillary electrophoresis technique for DNA analysis.

Citations

Dec 25, 2002·Journal of Chromatography. a·Julia Khandurina, András Guttman
Oct 16, 2003·Journal of Chromatography. a·Andras GuttmanXun Wang
Jan 27, 2011·Analytical Chemistry·Gwendolyn J AndersonRobert T Kennedy
Nov 2, 2007·Chemical Reviews·Karel Kleparník, Petr Bocek
May 30, 2002·Analytical Chemistry·Julia KhandurinaAndrás Guttman
Jul 15, 2005·Analytical Chemistry·Rongsheng LinMark A Burns
Apr 15, 2004·Analytical Chemistry·David G McLaren, David D Y Chen

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