Illumina HiSeq 2500 paired end sequencing; Chromothripsis 18 in multiple myeloma patient with rapid extramedullary relapse.
Chromothripsis 18 in multiple myeloma patient with rapid extramedullary relapse.
Female patient was diagnosed with MM in the Department of Internal Medicine – Hematooncology (IHOK), University Hospital Brno, in 11/2007. Patient's biological material was obtained after she signed the informed consent form approved by the Ethical committee of the hospital Plasma cells (CD138+) a were obtained from the bone marrow of multiple myeloma patients during routine clinical procedure. Plasma cells (CD138+) in mononuclear cell fraction were enriched by anti-CD138 immunomagnetic beads and sorted using magnetic activated cell sorting, MACS (AutoMACS, Miltenyi Biotec). Purity of CD138+ fraction was measured by flowcytometry.  Genomic DNA was extracted from sorted cells sing the Puregene Core kit according to manufacturer´s protocol. Extracted gDNA was diluted in TE buffer and stored in -20C. The capture probes are based on ultra-long RNA baits (120 nucleotides) with high sensitivity and specificity for the detection of SNPs and indels. Probe design captures all the coding regions of 42 genes including splicing sequences plus exonic hotspots of an additional 20 genes.
Experimental Factor: genotype
NRAS mutation c.181C>A; p.Gln61Lys