Comparison of fluorescent tag DNA labeling methods used for expression analysis by DNA microarrays
Comparison of fluorescent tag DNA labeling methods used for expression analysis by DNA microarrays
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DOI：
10.2144/02333rr05_11834a
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发表时间：
2002-09-01
期刊：
影响因子：
2.7
通讯作者：
Muckenthaler, M
中科院分区：
文献类型：
--
作者：
Richter, A;Schwager, C;Muckenthaler, M
DNA 微阵列的基因表达谱在生物医学研究的许多领域都有广泛的应用。该技术的协议尚未标准化，并且对于微阵列分析中的每个给定步骤，正在使用许多不同的协议。因此，不同实验室获得的结果可能难以比较。在这方面特别重要的是荧光 cDNA 探针的制备方法，该探针应定量反映待比较的两个样品中不同 mRNA 的丰度。在这里，我们系统地评估和比较了用于微阵列分析的荧光标记探针合成的五种不同的已发表和/或商业原理（直接标记、T7 RNA聚合酶扩增、氨基烯丙基标记、半抗原抗体酶标记和3-D多标记结构）。我们表明，单独的标记方法可以显着影响微阵列实验中获得的表达模式，并讨论每种方法各自的优点和局限性。
Gene expression profiling by DNA microarrays has found wide application in many fields of biomedical research. The protocols for this technique are not yet standardized, and for each given step in microarray analysis a number of different protocols are in use. As a consequence, results obtained in different laboratories can be difficult to compare. Of particular importance in this respect are the methods for the preparation of fluorescent cDNA probes that should quantitatively reflect the abundance of different mRNAs in the two samples to be compared. Here we systematically evaluate and compare five different published and/or commercial principles for the synthesis of fluorescently labeled probes for microarray analysis (direct labeling, T7 RNA polymerase amplification, arninoallyl labeling, hapten-antibody enzymatic labeling, and 3-D multi-labeled structures). We show that individual labeling methods can significantly influence the expression pattern obtained in a microarray experiment and discuss the respective benefits and limitations of each method.