TruSight Myeloid Sequencing Panel FAQs

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  • Protocol

  • Yes. The number of PCR cycles has been optimized for the number of amplicons per reaction. The recommended number of PCR cycles for different amplicon numbers is indicated in the user guide and ensures sufficient yield for the bead-based normalization while avoiding over-amplification.

    The ACD1 control is included to allow customers to assess the success of the library preparation. Illumina recommends that customers who are new to the assay include the ACD1 control. Technical Support will not be able to troubleshoot without this control. If the ACD1 control is included in the sequencing run, it should replace a test sample. For example, if the assay recommends eight samples per run and the ACD1 control is included, then there will be seven test samples plus the ACD1 control.

    One pair of oligos is designed for each amplicon. Hybridization of these oligos to unfragmented genomic DNA occurs in a 96‐well plate, followed by extension and ligation to form DNA templates consisting of the regions of interest flanked by universal primer sequences. Using indexed primers supplied with the kit, DNA templates are then PCR amplified, pooled into a single tube, and sequenced on the MiSeq System. For additional details on the assay, see the assay reference guide.