Pyrosequencing- A Pioneer Technique of New Epoch

Order of Publishing in Issue: 
10
Volume :10
Issue :3
July, 2016
Page No: 
290-298
Authors: 
Rekha Khandia and Ashok Munjal*
Address: 
Department of Genetics, Barkatullah University, Bhopal, 462026, India

Sanger’s dideoxy chain termination method was the widely used technology for DNA sequencing since its inception. However, the technique has its own limitations like it is costly, time consuming, labor intensive and faces the problem of clone biasness. Pyrosequencing has emerged as a novel sequencing technique, which offers potential advantages in terms of accuracy, cost effectiveness, flexibility, parallel processing, high throughput and further it can be easily automated. It can be widely accepted for the detailed characterization of nucleic acids. The technique is advantageous in DNA sequencing, genotyping, single nucleotide polymorphism analysis, allelic discrimination, allelic quantification and whole-genome sequencing. The technique may be broadly used in area of biotechnology, clinical genetics and pharmacogenetics. Due to efficacy and reliability, the technique may be used in the high-throughput analysis of bacterial and viral samples as well as metagenomic studies. Currently, the technique is the basis of next generation sequencing platform, offered by 454 Life Sciences as an array-based pyrosequencing technology.

Keywords: 
Pyrosequencing, real time PCR, diagnosis, luciferase, apyrase, emulsion PCR
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