What is pyrosequencing?

Posted July 22, 2020

Answer

Pyrosequencing is a DNA sequencing method based on the sequencing by synthesis principle. During the sequential addition of nucleotides onto a growing DNA strand, pyrophosphate (PPi) is released. A cascade of enzymatic reactions is then initiated by PPi, giving off chemiluminescent signals that can be detected and used to recognize the newly added nucleotide. One type of nucleotide is added and identified at a time, until full DNA molecule is sequenced.

Additional resources

Helixyte™ Green *10,000X Aqueous PCR Solution*

6-ROX glycine *25 uM fluorescence reference solution for PCR reactions*

Krishna, B. M., Khan, M. A., & Khan, S. T. (2019). Next-Generation Sequencing (NGS) Platforms: An Exciting Era of Genome Sequence Analysis. In Microbial Genomics in Sustainable Agroecosystems (pp. 89-109). Springer, Singapore.

Ju, J., Kim, D. H., Bi, L., Meng, Q., Bai, X., Li, Z., ... & Edwards, J. R. (2006). Four-color DNA sequencing by synthesis using cleavable fluorescent nucleotide reversible terminators. Proceedings of the National Academy of Sciences, 103(52), 19635-19640.