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Lab in a Tube: Purification, Amplification, and Detection of DNA Using Poly(2‐oxazoline) Multilayers
Author(s) -
Leiske Meike N.,
Hartlieb Matthias,
Paulenz Christian,
Pretzel David,
Hentschel Martin,
Englert Christoph,
Gottschaldt Michael,
Schubert Ulrich S.
Publication year - 2015
Publication title -
advanced functional materials
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 6.069
H-Index - 322
eISSN - 1616-3028
pISSN - 1616-301X
DOI - 10.1002/adfm.201404510
Subject(s) - materials science , polymer , monolayer , layer (electronics) , primer (cosmetics) , chemical engineering , contact angle , siloxane , polymer chemistry , nanotechnology , organic chemistry , chemistry , composite material , engineering
Fast and easy purification and amplification of DNA are prerequisites for the development of point‐of‐care diagnostics. For this reason covalent coatings of amine containing poly(2‐oxazoline)s (POx) on glass and poly(propylene) surfaces are prepared, to reversibly bind genetic material directly from biological samples. The polymer is deposited in a layer‐by‐layer process, whereas initial immobilization of macromolecules on the surface is accomplished by the use of an epoxy functionalized siloxane monolayer. Alternating treatment with polymer and cross‐linker leads to the construction of amine containing POx multilayers on the substrates. Successful deposition is investigated by confocal laser scanning microscopy (using labeled polymers), contact angle measurements, as well as reflectometric interference spectroscopy. The interaction of these layer systems with DNA regarding binding and temperature dependent release is studied using labeled genetic material. Finally, polymerase chain reaction (PCR) vessels are coated with POx layers on the inside, and used for quantitative real‐time PCR (qPCR) experiments. It is possible to bind genetic material directly from cell lysates to perform qPCR assays from surface adsorbed DNA within the same tube including amplification, as well as detection. The presented system displays an easy to use device for a point of care diagnostic.