EpiTect MSP Kit
For highly accurate methylation-specific PCR (MSP) without optimization
The EpiTect MSP Kit includes the novel genetically engineered HotStarTaq d-Tect Polymerase in a convenient master mix solution. Higher specificity of this polymerase produces reliable results. The polymerase efficiently discriminates between single-base mismatches at the 3' end of primers during annealing and extension. Since a single mismatch is sufficient for specific MSP results, the EpiTect MSP Kit even enables reliable MSP analysis of single CpG sites.
Novel genetically engineered polymerases, such as HotStarTaq d-Tect included in QIAGEN’s EpiTect MSP Kit, provide high PCR specificity (see figure "Sensitivity of HotStarTaq d-Tect"). This polymerase only extends primers that perfectly match the target DNA at the 3' primer end (the starting point of primer extension), enabling analysis of single CpG sites. This is in contrast to wild-type DNA polymerases, which often ignore 3' mismatches and extend mis-primed primers, producing nonspecific, false-positive methylation results (see figure "Increased mismatch discrimination").
The EpiTect MSP Kit enables simple development of new methylation-specific PCR assays. Primer design for MSP can be difficult, due to the need for each primer to cover several CpG sites. This has prevented the analysis of a single CpG site within a CpG island. However, since HotStarTaq d-Tect Polymerase only requires a single base mismatch at the 3' end of one primer, designing primers for MSP is simplified, and sensitive analysis of the methylation status of several and even individual CpG sites can be performed (see figure "Single base discrimination").
The EpiTect MSP Kit offers a master mix format, allowing convenient and easy reaction setup. HotStarTaq d-Tect Polymerase is provided in an inactive state, with no polymerase activity at ambient temperatures. This prevents the formation of misprimed products and primer dimers at low temperatures. HotStarTaq d-Tect Polymerase is activated by a 10-minute, 95°C incubation step, which can easily be incorporated into existing thermal cycling programs.
Standardized workflows in epigenetics
Accessing epigenetic information is of prime importance for many areas of biological and medical research — particularly oncology, but also stem cell research and developmental biology. However, the analysis of changes in DNA methylation is challenging due to the lack of standardized methods for providing reproducible data, particularly from limited sample material. With its newly introduced EpiTect solutions, QIAGEN makes available standardized, pre-analytical and analytical solutions: from DNA sample collection, stabilization and purification, to bisulfite conversion and real-time or end-point PCR methylation analysis or sequencing (see figure "Standardized workflows in epigenetics").
The EpiTect MSP Kit is dedicated for highly specific methylation analysis by end-point PCR.
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