研究目的
To develop and evaluate Morpholino Oligonucleotide crosslinked hydrogels as portable optical oligonucleotide biosensors for sensitive and selective detection of microRNA sequences, improving upon DNA-based systems in terms of thermal stability, salt independence, and sensitivity.
研究成果
MO crosslinked hydrogels demonstrate enhanced sensitivity, thermal stability, and salt independence compared to DNA-based systems, with a limit of detection as low as 10 pM. They are suitable for portable biosensing applications, as shown by mobile phone-based measurements, and hold promise for integration into various biosensor technologies.
研究不足
The study may have limitations in sensitivity at very low concentrations, potential for competitive displacement by non-target sequences, and the need for further optimization for real-world applications such as point-of-care diagnostics. The use of carbon nanopowder for contrast and specific equipment might not be universally accessible.
1：Experimental Design and Method Selection:
The study involved synthesizing MO crosslinked hydrogels via UV-initiated radical polymerization of acrylamide, functionalized MOs, and MBA. The design leveraged MO crosslinks for selective cleavage by target ssDNA sequences, inducing swelling measured optically.
2：Sample Selection and Data Sources:
Morpholino oligonucleotides and ssDNA sequences were purchased from GeneTools and IDT Technologies. Specific sequences (e.g., S1, B1, A1) were used, with miR-92a as a model analyte.
3：List of Experimental Equipment and Materials:
Materials included acrylamide, MBA, NaCl, HPK photoinitiator, carbon nanopowder, silicon oxide chips, and oligonucleotides. Equipment included a Dymax Bluewave 75 UV curing light source, Sony XCD-X710 Firewire Camera, MEDALight LP-300 lightbox, and a OnePlus 5t camera with a SODIAL(R) 30X Zoom LED Magnifier.
4：Experimental Procedures and Operational Workflow:
Pre-gel solutions were prepared, deposited on silanized chips, polymerized under UV light, washed, and swollen in buffer solutions with varying ssDNA concentrations, salt levels, and temperatures. Swelling was monitored optically using cameras and image analysis software.
5：Data Analysis Methods:
Gel volume changes were analyzed using custom MatLab code and Digimizer software, calculating percentage volume change and comparing responses to different sequences and conditions.
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