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Next Generation Sequencing for the Detection of Actionable Mutations in Solid and Liquid Tumors
Published on: September 20, 2016
Targeted Next-Generation Sequencing of Acute Leukemia
Eric Konnick1, Christina M Lockwood2, David Wu3
1Genetics and Solid Tumor Laboratory, Department of Laboratory Medicine, NW120, University of Washington, Box 357110, 1959 Pacific St., Seattle, WA, 98195, USA. Konnick@u.washington.edu.
Mutation profiling of acute leukemias aids in identifying key genetic mutations. A custom amplicon sequencing assay offers a customizable and efficient method for targeted resequencing in leukemia research.
Area of Science:
- Genetics
- Oncology
- Molecular Biology
Background:
- Mutation profiling is crucial for understanding acute leukemias, guiding prognosis, treatment, and diagnosis.
- Advancements in massively parallel sequencing enable cost-effective, simultaneous analysis of numerous genes.
Purpose of the Study:
- To describe the design and application of a TruSeq Custom Amplicon assay for acute leukemia gene mutation analysis.
- To present a customizable, amplicon-based targeted resequencing method for specific genomic regions.
Main Methods:
- Utilized a TruSeq Custom Amplicon assay for targeted resequencing.
- Designed the assay to interrogate genes critical in acute leukemias.
- Implemented a workflow supporting multiplexing of up to 1536 amplicons per reaction.
Main Results:
- The assay allows for customizable selection of genomic regions of interest.
- A single reaction can sequence up to 1536 amplicons, covering up to 650 kb.
- The workflow supports batch processing of up to 96 samples, depending on sequencing depth.
Conclusions:
- The described TruSeq Custom Amplicon assay provides a flexible and efficient approach for comprehensive mutation profiling in acute leukemias.
- This method facilitates targeted resequencing, enhancing the diagnostic and therapeutic potential of genetic analysis in leukemia.
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