Sequence specific colorimetric detection of DNA post-PCR using split DNAzyme

• Main Author: Aw, Darius Kang Lie
• Other Authors: Poh Chueh Loo
• Format: Final Year Project
• Language: English
• Published: 2010
• Subjects: DRNTU::Engineering::Chemical engineering::Biotechnology
• Online Access: http://hdl.handle.net/10356/39817
• Institution: Nanyang Technological University

The split G-quadruplex DNAzyme has become a valuable tool for visual detection. Most reports on its use have however been constrained to proof-of-concept demonstrations using synthetic oligonucleotides. This is due to the inherent complexities of the assay, and the multiple components involved. Herein, we have overcome several of these challenges and have successfully integrated asymmetric PCR with the visual detection step, allowing enriched targets from complex samples to be conveniently detected. This workflow brings the DNAzyme system closer to real-world detection applications. We have established the platform herein as a simple and robust method to determine the presence of target DNA sequences post-PCR, without the need for gels or other apparatus. Nanomolar concentrations of amplified targets were detected using the workflow established, enabling the detection of pathogenic targets and the discrimination of Single Nucleotide Polymorphism (SNP) targets through a color change reaction, observable just by eye.