Pyrosequencing for bacterial identification

Abstract

Sanger sequencing, first introduced in molecular laboratories 30 years ago, has been a useful tool in determining a vast array of DNA sequences. Over the years, modifications to the original technique have been made, solidifying Sanger sequencing as an efficient and robust technology. More recently, pyrosequencing has been developed as another technique for DNA sequencing. Pyrosequencing is a process of sequencing by synthesis which uses a substrate driven enzymatic reaction to detect sequential incorporation of nucleotides. Compared to traditional Sanger sequencing, this method provides a shorter sequence to analyze, offering faster turnaround times. There are many applications for pyrosequencing including sequence analysis, and SNP and CpG methylation detection and quantification. Here the sequence analysis application is used as an illustration of how pyrosequencing can be an effective approach for microbial identification.