Question

Compare and contract next generation sequencing.

1.Pyrosequencing

2.454 Technology

3.Illumina sequencing

plz give details

Answer 1

1.454 pyrosequencing technology is type od Pyrosequencing method which utilises the use of enzyme luciferase and ATP Sulfurylase, wher the genes are broken into nucleotides anf then joined with Adenosine-5- phosphosulfate to form ATP.This ATP is broken down into oxyluciferin, AMP and light in the presence of enzymes - luciferase and sufurylase. However, misinterpretation of homopolymers become the major disadvantage of this process.

While Illumina technology is the nexGen sequencing method that utilises a unique bridged amplification reaction occurring on the cell surface in such a way that each of the amplified DNA fragment produced from localised PCR get converted into clusters.These cluster are in the form of Single stick, which on addition of primers get bend into U-shape individually and the sequencing is then detected using fluorescent probe attached at 3'-OH of the strand. It is much cheaper than 454 pyrosequencing technology.