title: Expression analysis of multiple myeloma CD138 negative progenitor cells using single molecule microarray readout.
abstract: We present a highly sensitive bioanalytical microarray assay that enables the analysis of small genomic sample material. By combining an optimized cDNA purification step with single molecule cDNA detection on the microarray, the platform has improved sensitivity compared to conventional systems, allowing amplification-free determination of expression profiles with as little as 600ng total RNA. Total RNA from cells was reverse transcribed into fluorescently labeled cDNA and purified employing a precipitation method that minimizes loss of cDNA material. The microarray was scanned on a fluorescence chip-reader with single molecule sensitivity. Using the newly developed platform we were able to analyze the RNA expression profile of a subpopulation of rare multiple myeloma CD138 negative progenitor (MM CD138(neg)) cells. The high-sensitivity microarray approach led to the identification of a set of 20 genes differentially expressed in MM CD138(neg) cells. Our work demonstrates the applicability of a straight-forward single-molecule DNA array technology to current topics of molecular and cellular cancer research, which are otherwise difficult to address due to the limited amount of sample material.
authors: Biophysics Institute, Johannes Kepler University Linz, A-4040 Linz, Austria.
authors: Jaroslaw
authors: J
authors: Jacak
authors: Harald
authors: H
authors: Schnidar
authors: Leila
authors: L
authors: Muresan
authors: Gerhard
authors: G
authors: Regl
authors: Annemarie
authors: A
authors: Frischauf
authors: Fritz
authors: F
authors: Aberger
authors: Gerhard J
authors: GJ
authors: Schütz
authors: Jan
authors: J
authors: Hesse
mesh_terms: D045744:Cell Line, Tumor; D020869:Gene Expression Profiling / Q000379:methods*; D006801:Humans; D008856:Microscopy, Fluorescence / Q000379:methods*; D009101:Multiple Myeloma / Q000378:metabolism / Q000473:pathology*; D014411:Neoplastic Stem Cells / Q000737:chemistry / Q000166:cytology / Q000378:metabolism*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D013234:Stem Cells / Q000737:chemistry / Q000166:cytology / Q000378:metabolism*; D053668:Syndecan-1 / Q000235:genetics / Q000378:metabolism*
chemical_list: D012333:RNA, Messenger; D053668:Syndecan-1