Pyrosequencing is a type of DNA sequencing based on the “sequencing by synthesis” method. Unlike traditional Sanger sequencing, Pyrosequencing does not require fluorescence dye labeling or gel electrophoresis, while allowing quantitative, real time detection of sequencing events.
The Pyrosequencing platform is using the PSQ 96MA instrument to provide core services:
| Instrument | Vendors | Max Number of Read Per Run |
Read Length | Applications |
| PSQ 96MA | Biotage-Qiagen | 96 | 50-60bp | Quantitative DNA methylation analysis Low/Mid-throughput Genotyping |
Principle
Pyrosequencing technology allows real time detection of sequencing events using reliable chemistry and robust detection mechanisms. It uses an enzyme-cascade system, consisting of four enzymes and specific substrates to produce light upon nucleotide incorporation during DNA synthesis. The amount of light produced is proportional to the number of incorporated nucleotides. When light signal is detected, the base is registered and the next nucleotide is added. If the added nucleotide is not complementary to the next base in the template, no light will be generated.
Applications
- Quantitative DNA methylation analysis
- Mutation analysis and genotyping
- Allelic quantification
- Microbial identification and drug resistance typing
- Pharmacogenetics
- Clinical research
- Hematology research
