A versatile assay for high-throughput gene expression profiling on universal array matrices.
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.
Fan
Jian-Bing
JB
Illumina, Inc., San Diego, California 92121, USA. jfan@illumina.com
Yeakley
Joanne M
JM
Bibikova
Marina
M
Chudin
Eugene
E
Wickham
Eliza
E
Chen
Jing
J
Doucet
Dennis
D
Rigault
Philippe
P
Zhang
Baohong
B
Shen
Richard
R
McBride
Celeste
C
Li
Hai-Ri
HR
Fu
Xiang-Dong
XD
Oliphant
Arnold
A
Barker
David L
DL
Chee
Mark S
MS
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
D000728:Androgens; D018076:DNA, Complementary; D012313:RNA