Fan Jian-Bing, Yeakley Joanne M, Bibikova Marina, Chudin Eugene, Wickham Eliza, Chen Jing, Doucet Dennis, Rigault Philippe, Zhang Baohong, Shen Richard, McBride Celeste, Li Hai-Ri, Fu Xiang-Dong, Oliphant Arnold, Barker David L, Chee Mark S
Illumina, Inc., San Diego, California 92121, USA.
Genome Res. 2004 May;14(5):878-85. doi: 10.1101/gr.2167504.
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.
我们报告了一种灵活、灵敏且定量的基因表达谱分析系统,可用于同时检测96个样本中400多个基因,每个基因有三个探针。cDNA介导的退火、选择、延伸和连接(DASL)分析靶向特定转录本,使用含有可与通用阵列杂交的独特地址序列的寡核苷酸。使用该分析方法对激素处理的细胞系和激光捕获显微切割的癌组织进行检测,获得了细胞特异性基因表达谱。该分析方法得出的基因表达谱与通过qRT-PCR确定的结果一致。DASL分析已实现自动化,可与基于珠子的96阵列矩阵系统配合使用。这种高通量分析和读出系统准确高效,能够经济高效地分析数千个样本中数百个基因的表达情况。