A total of 40 elderly, transplant-ineligible patients with newly-diagnosed symptomatic MM staged according to the International Myeloma Working Group criteria 24 were prospectively studied after inclusion in the GEM2010MAS65 trial. In all patients, bone marrow (BM) aspirates were collected at diagnosis and after 9 cycles of induction therapy. In brief, patients treatment consisted of bortezomib, melphalan, prednisone (VMP) and lenalidomide plus low-dose dexamethasone (Rd), given in a sequential (n=21) or alternating (n=19) manner.
Extracted molecule
genomic DNA
Extraction protocol
Genomic DNA was digested with Nsp I and Sty I restriction enzymes, ligated to adapters with T4 DNA ligase and amplified. PCR products were concentrated and fragmented (DNAsal), and then end-labeled with deoxynucleotidil transferase
Label
biotin
Label protocol
Labelling and hybridizations were performed according to protocols from Affymetrix
Hybridization protocol
DNA was restriction digested, PCR amplified, fragmented, labeled and hybridized to each array according to the manufacturer's instructions.
Scan protocol
The Arrays were then washed using Affymetrix fluidics stations, and scanned using the Gene Chip Scanner 3000.
Data processing
ChAS console from Affymetrix was used to assess genome wide copy number frequencies Weighted Log2 Ratios from ChAS console
Phenotypic and genomic analysis of multiple myeloma minimal residual disease clonal plasma cells: a new model to understand chemoresistance [CytoScan750K Copy Number, LOH]
