Pyrosequencing is a DNA sequencing technique that detects pyrophosphate release upon nucleotide incorporation, allowing sequencing of short DNA sequences. It works by coupling the enzymatic activities of DNA polymerase, ATP sulfurylase, luciferase and apyrase to generate light signals proportional to incorporated nucleotides. The technique provides accurate, parallel sequencing that is easily automated compared to traditional Sanger sequencing, but is limited to smaller sequences due to non-linear light responses with longer homopolymers.

1. Pyrosequencing
1. Pyrosequencing Basics
2. Phyrosequencing Mechanism
3. Advantage/disvantage of Phyrosequencing

2. What is DNA Pyrosequencing?

3. Coupling Stage-(Polysulfurylase,Luciiferase)
Degradation Stage(Apyrase)
Catalyst (Adenosine phospho sulphate 5’ and Luciferin
What is DNA Pyrosequencing?

7. Pyrosequencing basics

8. Pyrosequencing Mechanism inside the well (glass
slide)

9. Advantage/disvantage of Phyrosequencing
1. Accurate
2. Parallel processing
3. Easily automated
4. Eliminate the needs for
labelled primers and
nucleotides
5. No need of gel
electrophoresis
1. Smaller sequences
2. Non linear lights
response after more
than 5-6 identical
nucleotides