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Lecture

Lab in a Tube: Or the Combination of DNA Purification and Amplification by Using a Poly(2 oxazoline) Based All-in-One System

Wednesday (10.05.2017)
12:30 - 12:50
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Since their discovery in the 1960’s, poly(2-oxazoline)s (POxs) have become an important polymer class for biomedical and bioanalytical applications.[1] By using the cationic ring-opening polymerization, it is possible to incorporate versatile functionalities into the polymer.[2] It has been shown that the polymerization of amino functionalized 2-oxazolines is possible when using Boc-protection groups.[3] The copolymerization with EtOx results in statistic copolymers that have characteristics, which can be easily fine-tuned by varying the monomer ratios. As a consequence, crosslinked structures, such as hydrogels can be prepared and used for successful DNA catch and release.[4]

In the present work, we want to show the establishment of a POx based DNA purification system, which is covalently surface bound to reaction vessels being used for the polymerase chain reaction (PCR).[5]

To achieve this aim, a POx bearing 20% amino functionalities was synthesized and covalently bound in a layer-by-layer approach to poly(propylene) (PP). A verification of this irreversible coating and the layer construction was performed by contact angle measurements, confocal laser scanning microscopy (CLSM), thermogravimetric analysis and RIFFS measurements.

With the aid of reversible DNA-binding studies by CLSM and RIFFS measurements, the optimum layer amount was determined and finally coated directly into PP PCR tubes. After optimization of the PCR protocol, bioanalytical investigations could be performed without elaborative sample preparation, paving the way for faster pathogen detection as an important part of point-of-care diagnostics.

[1] R. Hoogenboom, H. Schlaad, Polymers 2011, 3, 467-488.

[2] B. Guillerm, S. Monge, V. Lapinte, J.-J. Robin, Macromol. Rapid. Commun. 2012, 33, 1600-1612.

[3] M. Hartlieb, D. Pretzel, K. Kempe, C. Fritzsche, R. M. Paulus, M. Gottschaldt, U. S. Schubert, Soft Matter 2013, 9, 4693-4704.

[4] M. Hartlieb, D. Pretzel, C. Englert, M. Hentschel, K. Kempe, M. Gottschaldt, U. S. Schubert, Biomacromolecules 2014, 15, 1970-1978.

[5] M. N. Leiske, M. Hartlieb, C. Paulenz, D. Pretzel, M. Hentschel, C. Englert, M. Gottschaldt, U. S. Schubert, Adv. Func. Mater. 2015, 25, 2458-2466.

Speaker:
Meike Leiske
Friedrich Schiller University Jena
Additional Authors:
  • Dr. Matthias Hartlieb
    Friedrich Schiller University Jena
  • Christian Paulenz
    Analytik Jena AG
  • Dr. David Pretzel
    Friedrich Schiller University Jena
  • Martin Hentschel
    Analytik Jena AG
  • Christoph Englert
    Friedrich Schiller University Jena
  • Dr. Michael Gottschaldt
    Friedrich Schiller University Jena
  • Prof. Dr. Ulrich S. Schubert
    Friedrich Schiller University Jena