Core-Shell Beads as Microreactors for Phylogrouping of E. coli Strains
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Sreejith, Kamalalayam Rajan
Christie, Melody
Jin, Jing
Ooi, Chin Hong
Katouli, Mohammad
Stratton, Helen
Nguyen, Nam-Trung
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Abstract
Multiplex polymerase chain reaction (PCR) is an effective tool for simultaneous detection of target genes. Nevertheless, their use has been restricted due to the intrinsic interference between primer pairs. Performing several single PCRs in an array format instead of a multiplex PCR is a simple way to overcome this obstacle. However, there are still major technical challenges in designing a new generation of single PCR microreactors with a small sample volume, rapid thermal cycling, and no evaporation during amplification. We report a simple and robust core-shell bead array for a series of single amplifications. Four core-shell beads with a polymer coating and PCR mixture were synthesized using liquid marble formation and subsequent photo polymerization. Each bead can detect one target gene. We constructed a customised system for thermal cycling of these core-shell beads. Phylogrouping of the E. coli strains was carried out based on the fluorescent signal of the core-shell beads. This platform can be a promising alternative for multiplex nucleic acid analyses due to its simplicity and high throughput. The platform reported here also reduces the cycling time and avoids evaporation as well as contamination of the sample during the amplification process.
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Micromachines
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11
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8
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© 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
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Nanotechnology
PCR
core-shell bead
liquid marble
phylogrouping
simultaneous detection
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Gorgannezhad, L; Sreejith, KR; Christie, M; Jin, J; Ooi, CH; Katouli, M; Stratton, H; Nguyen, N-T, Core-Shell Beads as Microreactors for Phylogrouping of E. coli Strains., Micromachines, 2020, 11 (8), pp. 761-761