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首页> 外文期刊>Genomics >Efficacy of RNA amplification is dependent on sequence characteristics: implications for gene expression profiling using a cDNA microarray.
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Efficacy of RNA amplification is dependent on sequence characteristics: implications for gene expression profiling using a cDNA microarray.

机译：RNA扩增的效率取决于序列特征：使用cDNA微阵列进行基因表达谱分析的意义。

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Minute tissue samples or single cells increasingly provide the starting material for gene expression profiling, which often requires RNA amplification. Although much effort has been put into optimizing amplification protocols, the relative abundance of RNA templates in the amplified product is frequently biased. We applied a T7 polymerase-based technique to amplify RNA from two tissues of a cichlid fish and compared expression levels of unamplified and amplified RNA on a cDNA microarray. Amplification bias was generally minor and comprised features that were lost (1.3%) or gained (2.5%) through amplification and features that were scored as regulated before but unregulated after amplification (4.2%) or vice versa (19.5%). We examined 10 sequence-specific properties and found that GC content, folding energy, hairpin length and number, and lengths of poly(A) and poly(T) stretches significantly affected RNA amplification. We conclude that, if RNA amplification is used in gene expression studies, precedingexperiments controlling for amplification bias should be performed.
机译：微小的组织样本或单个细胞越来越多地提供基因表达谱分析的起始材料，而基因表达谱分析通常需要RNA扩增。尽管在优化扩增方案上已付出了很多努力，但扩增产物中RNA模板的相对丰度经常存在偏差。我们应用基于T7聚合酶的技术从丽鱼科鱼的两个组织中扩增RNA，并比较了cDNA微阵列上未扩增和扩增的RNA的表达水平。放大偏差一般较小，包括通过扩增丢失（1.3％）或获得（2.5％）的特征，以及在扩增之前被评定为受调控但在扩增后未被调节的特征（4.2％），反之亦然（19.5％）。我们检查了10种序列特异性的特性，发现GC含量，折叠能，发夹长度和数目以及poly（A）和poly（T）的长度显着影响RNA扩增。我们得出结论，如果在基因表达研究中使用RNA扩增，则应进行控制扩增偏差的先前实验。

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