Pyrosequencing and its application in biological science

Determining the DNA sequence is one of the most important techniques for identifying the sequence of nucleotides. In order to the remarkable advances in this field, pyrosequencing technique was created, has several applications in addition to determining the sequence of DNA. The use of this technique is in medical sciences and biotechnology, especially in the field of single nucleotide polymorphism, the identification of pathogenic and non-infectious microbes, DNA methylation, the identification of mutations in the major genes involved in the disease. This technique is based on synthesis and the enzyme system used in this method consists of four unique DNA Polymerase Klenow, ATP sulfurylase, Luciferase and Apyrase enzymes. Releasing of light produced by the enzyme luciferase determines the type of nucleotide present in the DNA chain, and the peaks of each nucleotide appear in the Pyrogram. The primer used in this method is flexible, and this unique feature has led to its high performance.Without using expensive materials, unlike other sequencing methods, is another advantage of this method.