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

초록

This work investigates the potential of GelMA hydrogels to serve as a supportive medium for PCR amplification, present-ing a promising substitute for traditional solution-based PCR methods. Liquid-phase PCR often faces issues like evapora-tion, handling errors, and contamination. Embedding PCR reagents in a hydrogel matrix offers a stable, enclosed environ-ment 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, HNMR, FTIR, XRD, UV-Vis. spectroscopy, and rheological analysis (loss and storage modulus) to confirm successful synthesis and tunable network for-mation. Amplification succeeded in uncrosslinked and UV-treated GeIMA 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.

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