Highlighted areas in the Pyrogram trace indicate variable CpG positions (light gray) and built-in bisulfite treatment controls (yellow). The methylation level of each CpG site is indicated in blue boxes on top of the Pyrogram trace. Data kindly provided by Dr. Triantafillos Liloglou, Roy Castle Lung Cancer Foundation, Molecular Oncology, Liverpool.
Apyrase, a nucleotide-degrading enzyme, continuously degrades unincorporated nucleotides and ATP. When degradation is complete, another nucleotide is added.
The figure illustrates the variation in methylation level between 5 different CpG sites in 4 individual tumor samples. Each sample was run in duplicate, and the concordance between the two samples clearly illustrates the reproducibility of Pyrosequencing technology.
The components of the PyroMark Q24 System are designed to make the Pyrosequencing workflow straightforward and efficient. Each step, from assay design to PCR amplification and preparation of sequencing templates, is supported by software, kits, reagents, and sample prep instrumentation optimized for Pyrosequencing.
Though small in size, the PyroMark Q24 manages all steps necessary to rapidly analyze up to 24 samples. Simply load your samples and reagents, upload your run file, and walk away. The PyroMark Q24 dispenses reagents and nucleotides to each well with precision. Light signals emitted are detected by 24 CCD sensors — one sensor per well — thereby eliminating signal crossover.
The PyroMark Q24 is designed as a stand-alone instrument, which makes it easy to place anywhere in the lab. Data are stored on the instrument hard drive and can be viewed on the instrument screen during a run. Additionally, all files are stored on the supplied USB stick, giving the user the flexibility to analyze data on any computer with PyroMark Q24 Software installed.
The PyroMark Q24 Vacuum Workstation enables conversion of PCR products into the single-stranded DNA for Pyrosequencing. Exposure of the PCR amplicons to a series of optimized solutions denatures and washes the DNA. This process is carried out for 24 samples in parallel and takes only a few minutes.
While the PyroMark Q24 combines ease-of-use, analysis versatility, and superior detection sensitivity, your workflow may demand higher throughput. The PyroMark Q96 ID and PyroMark Q96 MD accommodate this need by processing up to 96 samples in a single run. Furthermore, the automation options of the PyroMark Q96 MD enable hands-free processing of up to ten 96-well plates.