Targeted sequencing, in which only a selected set of genomic loci are sequenced, enables a much higher coverage of each target than what is obtained using whole genome or exome sequencing. Multiplex PCR offers a simple and affordable technique for specific capture of target regions and can be easily adapted to generate next-generation sequencing (NGS)-ready amplicons. Here we describe a multiplex PCR (MxPCR) approach for capturing 13 leukemia-associated mutation hotspots followed by MiSeq sequencing that enables robust detection of mutations with a variant allele fraction (VAF) as low as 0.8% (0.008) in blood DNA.
CITATION STYLE
Park, N., & Vassiliou, G. (2017). Design and application of multiplex PCR seq for the detection of somatic mutations associated with myeloid malignancies. In Methods in Molecular Biology (Vol. 1633, pp. 87–99). Humana Press Inc. https://doi.org/10.1007/978-1-4939-7142-8_6
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