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Pyrosequencing Background

Pyrosequencing Scientific Background Image Pyrosequencing technology, based on the principle of sequencing by real-time synthesis, provides both qualitative and quantitative genetic data.  Regardless of the complexity of the sequence, the simplicity of Pyrosequencing can provide quantifiable sequence data for an ~50 to 70 base-pair region of interest in one PCR-based assay.  More background information regarding the principle of Pyrosequencing technology can be found here.

Pyrosequencing can be applied to your research to:

  • Determine sequence variation, such as single nucleotide polymorphisms (SNPs), insertions/deletions of several bases, and short tandem repeats (STRs); quantify alleles; and determine heterozygosity
  • Quantify levels of DNA methylation at each CpG site of interest, as well as regional methylation levels at “CpG Islands”
  • Assay regions of sequence variation or DNA methylation that contain multiple sites of interest (e.g., multiple SNPs, multiple CpG sites, etc…)

Pyrosequencing can be useful for regions that have multiple sites and types of variation, where other PCR or hybridization-based assays may not allow for effective assay design.  However, if these sites of variation are greater than ~70 bases apart, other technologies such as Sanger Sequencing offered at the Laboratory of Molecular Technology may be more suitable.

For more information on this service and its applications to your research, visit our Pyrosequencing Protocols and Resources page.

Page last updated June 28, 2011 @ 5:56 pm