Sun Ye, Fan Wenhua, McCann Michael P, Golovlev Val
Sci-Tec, Inc., 10425 Cogdill Road, Suite 300, Knoxville, Tennessee 37932, USA.
Biotechnol Bioeng. 2009 Feb 15;102(3):960-4. doi: 10.1002/bit.22117.
The fabrication quality of microarrays significantly influences the accuracy and reproducibility of microarray experiments. In this report, we present a simple and fast quality control (QC) method for spotted oligonucleotide and cDNA microarrays. It employs a nonspecific electrostatic interaction of colloidal gold nanoparticles with the chemical groups of DNA molecules and other biomolecules immobilized on the microarray surface that bear positive or negative charges. An inexpensive flatbed scanner is used to visualize and quantify the binding of cationic gold particles to the anionic DNA probes on the microarray surface. An image analysis software was designed to assess the various parameters of the array spots including spot intensity, shape and array homogeneity, calculate the overall array quality score, and save the detailed array quality report in an Excel file. The gold staining technique is fast and sensitive. It can be completed in 10 min and detect less than 1% of the probe amount commonly recommended for microarrays. Compared to the current microarray QC method that utilizes the hybridization of probes with short random sequence oligonucleotides labeled with fluorophore, our gold staining method requires less time for the analysis, reduces the reagent cost, and eliminates the need for the expensive laser scanner.
微阵列的制作质量会显著影响微阵列实验的准确性和可重复性。在本报告中,我们提出了一种针对点阵寡核苷酸和cDNA微阵列的简单快速的质量控制(QC)方法。该方法利用胶体金纳米颗粒与固定在微阵列表面带有正电荷或负电荷的DNA分子及其他生物分子的化学基团之间的非特异性静电相互作用。使用一台价格低廉的平板扫描仪来可视化并定量阳离子金颗粒与微阵列表面阴离子DNA探针的结合情况。设计了一个图像分析软件来评估阵列点的各种参数,包括点强度、形状和阵列均匀性,计算阵列的整体质量得分,并将详细的阵列质量报告保存到Excel文件中。金染色技术快速且灵敏。它可在10分钟内完成,能检测到的探针量少于微阵列通常推荐量的1%。与当前利用探针与荧光团标记的短随机序列寡核苷酸杂交的微阵列QC方法相比,我们的金染色方法分析所需时间更少,降低了试剂成本,并且无需昂贵的激光扫描仪。