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- Conflict of Interest
- In relation to this article, we declare that there is no conflict of interest.
- Publication history
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Received August 5, 2025
Revised December 4, 2025
Accepted December 30, 2025
Available online March 25, 2026
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This is an Open-Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/bync/3.0) which permits
unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
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PCR Amplifi cation of HPRT1 Gene Within Photo-Crosslinkable Gelatin Methacrylate Hydrogel for Molecular Diagnostics
https://doi.org/10.1007/s11814-025-00636-5
Abstract
This work investigates the potential of GelMA hydrogels to serve as a supportive medium for PCR amplifi cation, 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 off ers a stable, enclosed environment
that helps overcome these challenges and supports effi cient amplifi cation. This study investigates the use of GelMA
hydrogels crosslinked with the lithium phenyl-2,4,6-trimethylbenzoylphosphinate (LAP) photo initiator for thermal PCR
amplifi cation of the HPRT1 gene. GelMA hydrogels were characterized by SEM, EDX, 1
HNMR, FTIR, XRD, UV–Vis.
spectroscopy, and rheological analysis (loss and storage modulus) to confi rm successful synthesis and tunable network formation.
Amplifi cation 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 amplifi cation, demonstrating reagent accessibility within the
hydrogel network. Increased LAP concentrations aff ected crosslink density and amplifi cation effi ciency. These results
demonstrate that GelMA hydrogels can serve as eff ective in-gel PCR platforms, combining enhanced stability with lower
contamination risk, holding promise for advanced biosensing and diagnostic applications.

