A multiplex microsphere bead assay for comparative RNA expression analysis using flow cytometry.
Comparative gene-expression profiling is an important tool in understanding molecular signatures of complex diseases as well as the responses of cells and tissues to external factors. With increasing microarray data, disease-specific molecular patterns are emerging but the acquisition of these data is expensive, difficult to customize and not well standardized. Once genome-wide scans identify differentially expressed genes in a given disease, cheaper, more easily customized methods will be needed for evaluating the expression of these genes in large population samples. Here we describe a novel multiplex microsphere bead assay (MBA) to compare gene expression levels. To test this assay we evaluated the expression levels of four transcripts (BRCA1, MGB1, DLG1 and ACT1) in normal and cancerous mammary tissue. The results were consistent with those generated by quantitative real-time PCR.
Fuja
Tannin
T
Developmental Biology Center, University of California Irvine, Irvine, CA 92697, USA. tfuja@uci.edu
Hou
Stephen
S
Bryant
Peter
P
D000199:Actins / Q000235:genetics / Q000378:metabolism; D019313:BRCA1 Protein / Q000235:genetics / Q000378:metabolism; D014408:Biomarkers, Tumor / Q000235:genetics / Q000378:metabolism*; D001940:Breast / Q000378:metabolism; D001943:Breast Neoplasms / Q000235:genetics / Q000378:metabolism*; D002478:Cells, Cultured; D016207:Cytokines / Q000235:genetics / Q000378:metabolism; D029721:Drosophila Proteins / Q000235:genetics / Q000378:metabolism; D005260:Female; D005434:Flow Cytometry / Q000379:methods*; D020869:Gene Expression Profiling / Q000379:methods*; D006801:Humans; D008863:Microspheres; D008875:Middle Aged; D015345:Oligonucleotide Probes; D012313:RNA / Q000032:analysis / Q000235:genetics* / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D014407:Tumor Cells, Cultured; D025521:Tumor Suppressor Proteins / Q000235:genetics / Q000378:metabolism
D000199:Actins; D019313:BRCA1 Protein; D014408:Biomarkers, Tumor; D016207:Cytokines; D029721:Drosophila Proteins; D015345:Oligonucleotide Probes; D025521:Tumor Suppressor Proteins; C070350:dlg1 protein, Drosophila; D012313:RNA