Skip to main content Accessibility help
×
Home

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

  • Stephen M. Lambert (a1), Jer-Kang Chen (a1), Claudia Chiesa (a1), George A. Fry (a1), Vergine C. Furniss (a1), Majid A. Mehrpouyan (a1) and Roger A. O'Neill (a1)...

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.

Copyright

References

Hide All
1. “Biomagnetic Techniques in Molecular Biology,” Technical Handbook, 2 ed., Dynal, New Hyde Park, NY, p. 90–96 (1995).
2. M.J., Gait, Oligonucleotide Synthesis.: A Practical Approach, IRL Press, Oxford, England, (1984).
3. S.S., Ghoshand G.F., Musso, Nucleic Acids Research, 15, p. 5353 (1987).
4. T.R., Gingeras, D.Y., Kwoh, and G.R., Davis, Nucleic Acids Research, 15, p. 5373 (1987).
5. F., Sanger, S., Micklen, and A.R., Coolson, Proc. Natl. Acad. Sci., 70, p. 5463 (1977).
6.J.M., Prober, et al., Science, 238, p. 336–41 (1987).

Related content

Powered by UNSILO

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

  • Stephen M. Lambert (a1), Jer-Kang Chen (a1), Claudia Chiesa (a1), George A. Fry (a1), Vergine C. Furniss (a1), Majid A. Mehrpouyan (a1) and Roger A. O'Neill (a1)...

Metrics

Full text views

Total number of HTML views: 0
Total number of PDF views: 0 *
Loading metrics...

Abstract views

Total abstract views: 0 *
Loading metrics...

* Views captured on Cambridge Core between <date>. This data will be updated every 24 hours.

Usage data cannot currently be displayed.