Principles of Pyrosequencing
Applications of Pyrosequencing
Global DNA methylation
Luminometric Methylation Assay (LUMA) is a high throughput and quantitative method to estimate genome-wide DNA methylation. The starting material is genomic DNA, and the method uses combined DNA cleavage by methylation-sensitive enzymes and polymerase extension assay using Pyrosequencing. The assay can be completed in ~50 mins (excluding the digestion process).
Gene-specific DNA methylation
The Pyrosequencer can be used to detect gene-specific DNA methylation patterns at specific genomic region (e.g., promoter of gene of interest). The average length size to be tested is ~100 bp. The starting material is bisulfite-treated genomic DNA. The assay can be completed in ~ 60 mins (excluding the PCR preparation).