Genetic Heterogeneity and Drug Resistance Mechanisms of Relapsed Refractory Multiple Myeloma: Sample SI_17828
"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). RNA-seq was performed by an exome-capture transcriptome platform. Libraries were prepared using 1-2 ug of total RNA. cDNA synthesis, end-repair, A-base addition, and ligation of the Illumina index adapters were performed according to Illumina's TruSeq RNA protocol (Illumina). Libraries were size-selected on 3% agarose gel. Recovered fragments were enriched by PCR using Phusion DNA polymerase (New England Biolabs) and purified using AMPure XP beads (Beckman Coulter). Capture transcriptomes were captured by Agilent SureSelect Human All Exon v.4 probes following the manufacturer's protocol. Library quality was measured on an Agilent 2100 Bioanalyzer for product size and concentration."