For analyzing gene-specific sequence variation or methylation on PyroMark Q96 ID or PyroMark Q96 MD
Detection and quantification of mutations in cancer-related genes
Quantification of individual or multiple CpG sites in different genes
Estimation of global CpG methylation in defined regions of interest
Sequence context provides built-in quality control of the assays
PyroMark Q96 Tests are a range of predesigned assays suitable for highly reliable, quantitative analysis of mutations or SNPs, as well as CpG methylation. Tests designated specifically with 'Q96' are optimized for use with the PyroMark Q96 MD and/or PyroMark Q96 ID Instruments. Other tests, such as PyroMark HFE and PyroMark MTHFR, can additionally be performed on the PyroMark Q24 (with the exception of PyroMark APOE).
For quantification of global methylation level in transposable elements; only for use with PyroMark Q96 MD
PyroMark APOE, PyroMark HFE, PyroMark MTHFR, PyroMark Q96 CpG LINE-1, PyroMark Q96 CpG MGMT, PyroMark Q96 CpG MLH1, PyroMark Q96 CpG p16, and PyroMark Q96 CpG Prader-Willi tests are intended for molecular biology applications. These products are not intended for the diagnosis, prevention, or treatment of a disease.
PyroMark p16 analysis.
[A] A normal blood DNA sample and [B] a DNA sample isolated from a colon tumor were analyzed. Blue columns indicate the measured CpG sites, and the individually measured degree of methylation at each site. Yellow columns indicate QC for completion of the bisulfite treatment.
The PyroMark Q96 Tests are highly suitable for accurately measuring the CpG methylation status, as exemplified with the PyroMark Q96 CpG p16 Test (see figure PyroMark p16 analysis).
PyroMark Q96 Tests used in combination with PyroMark Q96 MD and/or PyroMark Q96 ID Systems enable accurate, quantitative mutation (or SNP) and methylation analysis by Pyrosequencing. The assays consist of PCR primers and a sequencing primer.
Prior to Pyrosequencing, a biotinylated PCR product is generated. Biotinylated PCR products are bound to streptavidin-coated Sepharose beads. The beads are then captured with the vacuum tool on the PyroMark Vacuum Workstation, and throroughly washed, generating single-stranded DNA suitable for sequencing. PyroMark Q96 Tests are predesigned assays for methylation analysis by Pyrosequencing. Each test includes the primers necessary for generating the biotinylated PCR product, as well as the primer for the Pyrosequencing reaction.
PyroMark Q96 Tests enable highly accurate, quantitative methylation and mutation (or SNP) analysis applicable in several fields of research, including cancer and epigenetics.
For 800 reactions: 2x PyroMark PCR Master Mix (includes HotStarTaq DNA Polymerase and optimized PyroMark Reaction Buffer containing 3 mM MgCl2 and dNTPs), 10x CoralLoad Concentrate, 5x Q-Solution, 25 mM MgCl2, and RNase-Free Water