Front-End Processing of Cell Lysates for Enhanced Chip-Based Detection
Final rept. 1 Mar 2003-28 Feb 2006
CARNEGIE-MELLON UNIV PITTSBURGH PA
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The goal of this project was to develop a means to separate target DNA oligomers from complex mixtures using a tag-and separate approach that involves the use of surfactants that bind specifically to short sequences of the DNA targets. The surfactants include a peptide nucleic acid PNA segment to provide highly specific binding to DNA targets. Separation is achieved using capillary electrophoresis, the standard method of fluid manipulation used in lab-on-a-chip devices. A small unknown sample is first mixed with the PNA surfactants PNAA to tag the DNA targets, and then the sample is flushed with conventional surfactant micelles to pick up the tagged DNA targets. The method has several advantages over other specific DNA separation methods, such as the use of magnetic beads. Binding of PNAA to DNA occurs in solution, rather than on a bead surface, improving binding kinetics and preventing fouling of the bead surface by adsorption of proteins or lipids. The method is also compatible with longer DNA targets possessing overhanging stretches of DNA. The use of surfactants also helps to keep the capillary wall clean and free of adsorbed protein and lipid. The method can also distinguish both sequence and molecular weight of the target DNA.
- Physical Chemistry
- Electrical and Electronic Equipment
- Atomic and Molecular Physics and Spectroscopy