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February 10, 2012Journal of Molecular DiagnosticsOpen Access

A Virtual Pyrogram Generator to Resolve Complex Pyrosequencing Results

Guoli Chen Matthew T. Olson Alan M. O’Neill Alexis L. Norris Katie Beierl Shuko Harada Marija Debeljak Keila Rivera‐Román Samantha Finley Amanda Stafford Christopher D. Gocke Ming‐Tseh Lin James R. Eshleman

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Key Points

To develop and validate Pyromaker, a software that generates simulated pyrograms to analyze complex pyrosequencing results.
Developed Pyromaker to simulate pyrograms based on user inputs and hypothesized mutations.
Validated simulations against actual pyrograms for mutations in KRAS, BRAF, GNAS, and p53 genes.
Utilized two analytical strategies: hypothesis-based matching and iterative reconstruction for mutation identification.
All 18 single-base mutations in KRAS codons 12 and 13 produced unique pyrograms; complex mutations also generated distinct traces.
Pyromaker successfully identified complex mutations, confirmed by cloning and sequencing methods.
Combining pyrosequencing with Pyromaker produced the most efficient mutation identification strategy.

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Implication

Software generates simulated pyrograms for mutation detection in KRAS and related genes, aiding analysis.

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Cite This Study

Chen et al. (2012) studied this question.

synapsesocial.com/papers/6a214180a2a97f3a085addcf https://doi.org/10.1016/j.jmoldx.2011.12.001

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