Genetic Heterogeneity and Drug Resistance Mechanisms of Relapsed Refractory Multiple Myeloma: Sample SI_21003
"Tissues with the highest tumor content for each case were disrupted by 5 mm beads on a Tissuelyser II (Qiagen). Tumor genomic DNA and total RNA were purified from the same sample using an AllPrep DNA/RNA/miRNA kit (Qiagen). RNA integrity was measured on an Agilent 2100 Bioanalyzer using RNA Nano reagents (Agilent Technologies). To prepare exome libraries, 1-3 ug of genomic DNA was sheared using a Covaris S2 to a peak target size of 250 base pairs (bp). Fragmented DNA was concentrated using AMPure beads, followed by end-repair, A-base addition, ligation of the Illumina indexed adapters, and size selection on 3% Nusieve agarose gels (Lonza). Fragments between 300 to 350 bp were recovered, amplified using Illumina index primers, and purified by AMPure beads. One microgram of the library was hybridized to the Agilent SureSelect Human All Exon v.4. The targeted exon fragments were captured and enriched following the manufacturer's protocol (Agilent)."