title: A versatile assay for high-throughput gene expression profiling on universal array matrices.
abstract: We report a flexible, sensitive, and quantitative gene-expression profiling system for assaying more than 400 genes, with three probes per gene, for 96 samples in parallel. The cDNA-mediated annealing, selection, extension and ligation (DASL) assay targets specific transcripts, using oligonucleotides containing unique address sequences that can hybridize to universal arrays. Cell-specific gene expression profiles were obtained using this assay for hormone-treated cell lines and laser-capture microdissected cancer tissues. Gene expression profiles derived from this assay were consistent with those determined by qRT-PCR. The DASL assay has been automated for use with a bead-based 96-array matrix system. The combined high-throughput assay and readout system is accurate and efficient, and can cost-effectively profile the expression of hundreds of genes in thousands of samples.
authors.affiliation: Illumina, Inc., San Diego, California 92121, USA. jfan@illumina.com
authors.forename: Jian-Bing
authors.initials: JB
authors.lastname: Fan
authors.forename: Joanne M
authors.initials: JM
authors.lastname: Yeakley
authors.forename: Marina
authors.initials: M
authors.lastname: Bibikova
authors.forename: Eugene
authors.initials: E
authors.lastname: Chudin
authors.forename: Eliza
authors.initials: E
authors.lastname: Wickham
authors.forename: Jing
authors.initials: J
authors.lastname: Chen
authors.forename: Dennis
authors.initials: D
authors.lastname: Doucet
authors.forename: Philippe
authors.initials: P
authors.lastname: Rigault
authors.forename: Baohong
authors.initials: B
authors.lastname: Zhang
authors.forename: Richard
authors.initials: R
authors.lastname: Shen
authors.forename: Celeste
authors.initials: C
authors.lastname: McBride
authors.forename: Hai-Ri
authors.initials: HR
authors.lastname: Li
authors.forename: Xiang-Dong
authors.initials: XD
authors.lastname: Fu
authors.forename: Arnold
authors.initials: A
authors.lastname: Oliphant
authors.forename: David L
authors.initials: DL
authors.lastname: Barker
authors.forename: Mark S
authors.initials: MS
authors.lastname: Chee
mesh_terms: D000728:Androgens / Q000494:pharmacology; D000818:Animals; D002460:Cell Line; D045744:Cell Line, Tumor; D018076:DNA, Complementary / Q000096:biosynthesis / Q000235:genetics; D020869:Gene Expression Profiling / Q000379:methods*; D005786:Gene Expression Regulation / Q000187:drug effects / Q000235:genetics; D006801:Humans; D007834:Lasers; D008297:Male; D051379:Mice; D042282:Microdissection / Q000379:methods; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D009928:Organ Specificity / Q000187:drug effects / Q000941:ethics / Q000235:genetics; D011471:Prostatic Neoplasms / Q000378:metabolism / Q000473:pathology; D012313:RNA / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D012680:Sensitivity and Specificity
chemical_list: D000728:Androgens; D018076:DNA, Complementary; D012313:RNA