Home Pyrosequencing Technology Overview

Technology Overview

Print
PSQ 96MA Pyrosequencer

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.

The principle of pyrosequencing

 

Applications

  • Quantitative DNA methylation analysis
  • Mutation analysis and genotyping
  • Allelic quantification
  • Microbial identification and drug resistance typing
  • Pharmacogenetics
  • Clinical research
  • Hematology research