Pyrosequencing for SNP genotyping.

摘要: Pyrosequencing is a real-time DNA sequencing method. It based on the transformation of pyrophosphates, released during elongation by polymerase, into measurable light. During elongation, single pyrophosphate molecule following incorporation nucleotide. In pyrosequencing reaction, pyrophosphates are then rapidly converted sulfurylase to adenosine triphosphate, which in turn utilized luciferase produce Within standardized conditions, this reaction accomplished few milliseconds and light produced can be registered with CCD camera. Therefore, it becomes possible quantitatively measure nucleotides incorporated. This approach has been automated different platforms used for wide variety applications, such as single-nucleotide polymorphism (SNP) genotyping, sequencing, loss heterozygosity analysis, CpG methylation studies. Here we describe entire process, focusing our attention SNP giving examples some other applications.