Wu Haoxiang, Bynum James, Stavchansky Salomon, Bowman Phillip
College of Pharmacy, University of Texas at Austin, Austin, USA.
Biotechniques. 2008 Nov;45(5):573-5. doi: 10.2144/000112952.
DNA microarrays are powerful tools for global analysis of gene transcript expression. However, their high cost and the need for replication have limited their use. Here, we report a new stripping technique applicable to microarrays hybridized with cRNA with RNase H that is reproducible, leaving the DNA oligonucleotide probes intact and available for adding two additional uses. A Pearson correlation was used to assess the agreement between the first-round hybridization and the second- and third-round hybridizations. Significant correlations (R2, 0.9893 and 0.975; P < 0.001) were observed among virgin arrays and stripped arrays hybridized with the same sample. Additionally, statistical class comparison analysis globally indicated that there were essentially no differences detected following three hybridizations. Dye-swapped microarrays produced similar results. However, arrays stripped with RNase H exhibited decreased efficiency of hybridization signal with increasing use. In the present study, the oligonucleotide microarrays can be used three times.
DNA微阵列是用于基因转录本表达全局分析的强大工具。然而，其高昂的成本和对重复实验的需求限制了它们的应用。在此，我们报告了一种新的洗脱技术，适用于与用RNase H处理的cRNA杂交的微阵列，该技术具有可重复性，能使DNA寡核苷酸探针保持完整并可供额外进行两次使用。使用Pearson相关性来评估第一轮杂交与第二轮和第三轮杂交之间的一致性。在与相同样品杂交的原始阵列和洗脱后的阵列之间观察到显著的相关性（R2分别为0.9893和0.975；P < 0.001）。此外，统计类别比较分析总体表明，经过三次杂交后基本未检测到差异。染料交换微阵列产生了相似的结果。然而，随着使用次数增加，用RNase H洗脱的阵列杂交信号效率降低。在本研究中，寡核苷酸微阵列可使用三次。
