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<p>This work investigates the potential of GelMA hydrogels to serve as a supportive medium for PCR amplification, presenting a promising substitute for traditional solution-based PCR methods. Liquid-phase PCR often faces issues like evaporation, handling errors, and contamination. Embedding PCR reagents in a hydrogel matrix offers a stable, enclosed environment that helps overcome these challenges and supports efficient amplification. This study investigates the use of GelMA hydrogels crosslinked with the lithium phenyl-2,4,6-trimethylbenzoylphosphinate (LAP) photo initiator for thermal PCR amplification of the HPRT1 gene. GelMA hydrogels were characterized by SEM, EDX, <sup>1</sup>HNMR, FTIR, XRD, UV–Vis. spectroscopy, and rheological analysis (loss and storage modulus) to confirm successful synthesis and tunable network formation. Amplification succeeded in uncrosslinked and UV-treated GelMA without LAP but failed when PCR components were present during UV-induced crosslinking with LAP, which may be due to radical-induced DNA or enzyme damage. Notably, loading PCR reagents after crosslinking restored amplification, demonstrating reagent accessibility within the hydrogel network. Increased LAP concentrations affected crosslink density and amplification efficiency. These results demonstrate that GelMA hydrogels can serve as effective in-gel PCR platforms, combining enhanced stability with lower contamination risk, holding promise for advanced biosensing and diagnostic applications.</p>

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PCR Amplification of HPRT1 Gene Within Photo-Crosslinkable Gelatin Methacrylate Hydrogel for Molecular Diagnostics

  • 期刊论文

  • 发表日期：

    2026年1月13日

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  • Hyerin Nam,
  • Ajith Mohanasundaran,
  • Jeongha Yoo,
  • Hansol Lee,
  • Jongsung Kim

摘要

This work investigates the potential of GelMA hydrogels to serve as a supportive medium for PCR amplification, presenting a promising substitute for traditional solution-based PCR methods. Liquid-phase PCR often faces issues like evaporation, handling errors, and contamination. Embedding PCR reagents in a hydrogel matrix offers a stable, enclosed environment that helps overcome these challenges and supports efficient amplification. This study investigates the use of GelMA hydrogels crosslinked with the lithium phenyl-2,4,6-trimethylbenzoylphosphinate (LAP) photo initiator for thermal PCR amplification of the HPRT1 gene. GelMA hydrogels were characterized by SEM, EDX, 1HNMR, FTIR, XRD, UV–Vis. spectroscopy, and rheological analysis (loss and storage modulus) to confirm successful synthesis and tunable network formation. Amplification succeeded in uncrosslinked and UV-treated GelMA without LAP but failed when PCR components were present during UV-induced crosslinking with LAP, which may be due to radical-induced DNA or enzyme damage. Notably, loading PCR reagents after crosslinking restored amplification, demonstrating reagent accessibility within the hydrogel network. Increased LAP concentrations affected crosslink density and amplification efficiency. These results demonstrate that GelMA hydrogels can serve as effective in-gel PCR platforms, combining enhanced stability with lower contamination risk, holding promise for advanced biosensing and diagnostic applications.