A new web-based application has been introduced to help researchers more accurately define complex genetic mutations that are hard to identify with Sanger sequencing and pyrosequencing.

Pyromaker produces simulated programs based on user input, and could aid in the diagnosis and selection of treatment for cancer. It can interpret complex results based on the data inputted, including the percentage of tumour and normal cells, the wild-type sequence, the dispensation order, and any number of mutant sequences.

The Pyromaker makes a virtual trace of the expected program by calculating the relative mutant and wild-type allele percentages and using the results to generate the expected signal at each point in the dispensation sequence.

Senior author James R. Eshleman, MD, PhD, of Johns Hopkins University School of Medicine, explains: "Pyromaker can quickly and efficiently test the possibilities that can explain a complicated polysequencing result.

"Additional methods, such as Pyromaker analysis or TA cloning and sequencing, allow one to definitively diagnose the variant allele.”

Lab Asia Dec 2025

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