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Conjugation of DNA to Plasma-Treated Polymer Surfaces and Their Application to Multiplex DNA Sequencing

Published online by Cambridge University Press:  10 February 2011

Stephen M. Lambert
Affiliation:
PE Applied Biosystems, 850 Lincoln Centre Drive, Foster City, CA 94404 USA
Jer-Kang Chen
Affiliation:
PE Applied Biosystems, 850 Lincoln Centre Drive, Foster City, CA 94404 USA
Claudia Chiesa
Affiliation:
PE Applied Biosystems, 850 Lincoln Centre Drive, Foster City, CA 94404 USA
George A. Fry
Affiliation:
PE Applied Biosystems, 850 Lincoln Centre Drive, Foster City, CA 94404 USA
Vergine C. Furniss
Affiliation:
PE Applied Biosystems, 850 Lincoln Centre Drive, Foster City, CA 94404 USA
Majid A. Mehrpouyan
Affiliation:
PE Applied Biosystems, 850 Lincoln Centre Drive, Foster City, CA 94404 USA
Roger A. O'Neill
Affiliation:
PE Applied Biosystems, 850 Lincoln Centre Drive, Foster City, CA 94404 USA
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Abstract

Plasma treatment and deposition techniques have been used to facilitate the covalent attachment of DNA to polymer surfaces. The variety of surface functional groups that can be created by plasma techniques enables different chemical conjugation routes to be explored. For example, a phosphodiester linkage can be used for surfaces with hydroxyl groups. Likewise, a carboxyamide linkage can be used for surfaces with carboxyl or amine groups. The primary application of the engineered materials has been the hybridization-based separation of multiplexed DNA sequencing products. Traditionally, sequencing reactions are performed individually on single templates. Multiplex sequencing offers reagent and time savings by permitting multiple sequencing reactions on single or multiple templates. The method developed uses recoverable DNA sequencing primers with additional “capture sequences” attached to the 5' end. The capture sequences are designed to be complementary to “binding sequences” covalently attached to the plasma-treated polymer supports. When a solution of the extended recoverable primers is exposed to a polymer support, primers with complementary capture sequences hybridize to the immobilized binding sequences. Contacting a multiplexed sample with a series of solid supports (each having a different binding sequence) selectively removes each set of sequencing products from the mixture. Washing each solid support, followed by releasing the hybridized DNA, results in isolated and purified sequencing products that are amenable to analysis by gel or capillary electrophoresis.

Type
Research Article
Copyright
Copyright © Materials Research Society 1999

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