Pyrosequencing

Pyrosequencing is a method of sequencing that relies on light emitted in an enzymatic reaction set in motion by the release of pyrophosphate when a base is added during the sequencing process. The results are quantitative, making the method ideal for sequencing samples of known sequence that may have mutations in proportions as low as 5%, such as tumor samples or mixed viral populations.

This method can also be used for characterization of contiguous and multivariable mutations, LOH analysis, unambiguous, fully quantitative genotyping that distinguished multi-site variations from single nucleotide polymorphisms.

Microbial identification and drug resistance typing are also potential uses of this method. The system provides rapid and reliable high-throughput screening. Mutations can be accurately identified without use of expensive labels and dyes.

Specific regions of methylation can be assayed via sequencing on the Pyromark Q24 system as well. Bisulfite converted DNA is PCR amplified with a biotinylated primer pair. The non-biotinylated strand is sequenced in the Pyromark system.

• Qiagen has over 30,000 assays designed for CpG Islands throughout the human genome, which can be ordered through our Geneglobe.com service.
• Custom regions can be amplified; we are happy to assist with custom assay design, using the Pyromark Assay Design Software package.  This is best done via screen sharing.  You provide the specific CpG or SNP of interest, plus 200 bp of flanking sequence on each side, and then we will work through the possible assay designs together. Please email Roxann Ashworth to arrange a time.