Hydrogels for Efficient Multiplex PCR
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pISSN 1226-8372 eISSN 1976-3816
REVIEW PAPER
Hydrogels for Efficient Multiplex PCR Juny Shin and Cheulhee Jung
Received: 2 May 2020 / Revised: 23 May 2020 / Accepted: 23 May 2020 © The Korean Society for Biotechnology and Bioengineering and Springer 2020
Abstract Multiplex PCR simultaneously detects several different DNA targets in a given sample and has, therefore, gained considerable interest for use in examining a genetic target whose number rapidly increases in a single phenotype. Conventional real-time PCR lacks sufficient multiplex capacity, as the limited number of color channels restricts the number of simultaneously detectable targets to six. On the contrary, DNA-based hydrogel microparticles immobilized with primers provide outstanding multiplex capacity, as they can encode for various patterns with a level of coding capacity high enough to identify ≥ 105 targets. Moreover, hydrogel microparticles comprising polyethylene glycol can be made extremely porous, as it is easy to control their porosity and hydrophilic properties. Their high compatibility for aqueous biochemical reactions also ensures an environment and efficiency similar to amplification reactions occurring in aqueous media. The applicability of DNAbased encoded hydrogel microparticles with fixed primers has been widely proven through various hydrogel particlebased bioassays. This review focuses on the advancements in diverse approaches for an efficient multiplex PCR based on such microparticles. Herein, multiple strategies for constructing and encoding a “hydrogel” are described. Moreover, various methods for an efficient hydrogel PCR and the latest developments regarding multiple forms of bioassays that utilize hydrogel PCR are illustrated using selected examples. Lastly, this review provides insights into the prospects for encoded hydrogel microparticles and the challenges for their application. Keywords: hydrogel, multiplex PCR, encoded hydrogel, PEGDA, primer conjugation Juny Shin, Cheulhee Jung* Department of Biotechnology, College of Life Sciences and Biotechnology, Korea University, Seoul 02841, Korea Tel: 82-2-3290-3023 E-mail: [email protected]
1. Introduction Hydrogels are cross-linked hydrophilic polymers that exhibit high water swellability. Hence, they can contain a large quantity of water and exhibit a level of flexibility similar to that of natural tissues or aqueous environments [1]. Hydrogels have characteristics such as excellent adjustability and high flexibility and biocompatibility that provide the potential for application in a variety of fields, including bioassay diagnostics [2], drug delivery [3], cancer therapy [4], and tissue engineering [5]. Since hydrogels rely on the properties of their constituent polymers, their reactions may be sensitive to various physical and chemical changes, such as variations in pH [6-9], ionic strength [10-12], electric field [13-15], heat [16-18], ultrasound irradiation [19,20], and magnetic stimulation [21,22], which limits their applications [23]. The introduction of nucleotides
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