The QIAact Myeloid DNA UMI Panel is provided as two primer mix tubes, with approximately 400 primers per tube. The QIAact Myeloid DNA UMI Panel is designed to enrich specific target regions in select genes (ASXL1, CALR, CBL, CEBPα, CSF3R, DNMT3A, EZH2, FLT3, IDH1, IDH2, JAK2, KIT, KRAS, MPL, NPM1, NRAS, RUNX1, SETBP1, SF3B1, SH2B3, SRSF2, TET2, TP53, U2AF1, ZRSR2) using 40 ng of DNA.
Genomic DNA samples are first fragmented, end-repaired and A-tailed using a single, controlled multi-enzyme reaction. The prepared DNA fragments are then ligated at their 5’ ends to a GeneReader specific adapter containing a unique molecular index (UMI) and a nine (9) base-pair (bp) sample-specific bar code.
Ligated DNA molecules are subject to limited cycles of target enrichment PCR, with one gene-specific primer targeting a region and one universal forward primer complimentary to an adapter sequence. This reaction ensures that intended targets and UMIs are enriched sufficiently to be represented in the final library. A universal PCR with GeneReader specific sequences is then carried out to amplify the targets and complete the library.
Once the library is sequenced, results can be analyzed using the QIAact Myeloid DNA UMI Panel workflow, which will automatically perform all steps necessary to generate a DNA sequence variant report from your raw NGS data. All detected variants can be further interpreted by QIAGEN Clinical Insight (QCI) analysis.