Sequence specific colorimetric detection of DNA post-PCR using split DNAzyme
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. H...
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| Format: | Final Year Project |
| Language: | English |
| Published: |
2010
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| Subjects: | |
| Online Access: | http://hdl.handle.net/10356/39817 |
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| Institution: | Nanyang Technological University |
| Language: | English |
| Summary: | 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. |
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