Department of Molecular Pharmacology, Beckman Research Institute of the City of Hope, Duarte, California 91010-3000, USA.
RNA. 2011 Dec;17(12):2256-62. doi: 10.1261/rna.028621.111. Epub 2011 Oct 20.
Small RNAs (smRNAs) encompass several different classes of short noncoding RNAs. Progress in smRNA research and applications has coincided with the advance of techniques to detect them. Next-generation sequencing technologies are becoming the preferred smRNA profiling method because of their high-throughput capacity and digitized results. In our small RNA profiling study using Solexa, we observed serious biases introduced by the 5' adaptors in small RNA species coverage and abundance; therefore, the results cannot reveal the accurate composition of the small RNAome. We found that the profiling results can be significantly optimized by using an index pool of 64 customized 5' adaptors. This pool of 64 adaptors can be further reduced to four smaller index pools, each containing 16 adaptors, to minimize profiling bias and facilitate multiplexing. It is plausible that this type of bias exists in other deep-sequencing technologies, and adaptor pooling could be an easy work-around solution to reveal the "true" small RNAome.
小 RNA（smRNA）包含几种不同类别的短非编码 RNA。随着检测技术的进步，smRNA 的研究和应用也取得了进展。下一代测序技术因其高通量能力和数字化结果而成为首选的 smRNA 分析方法。在我们使用 Solexa 进行的小 RNA 分析研究中，我们观察到小 RNA 种类覆盖和丰度的 5' 接头带来了严重的偏差；因此，结果无法揭示小 RNA 组的准确组成。我们发现，通过使用 64 个定制的 5' 接头的索引池，可以显著优化分析结果。该 64 个接头池可以进一步减少到四个较小的索引池，每个池包含 16 个接头，以最大限度地减少分析偏差并促进多重分析。在其他深度测序技术中可能存在这种类型的偏差，而接头池可能是一个简单的解决方法，可以揭示“真实”的小 RNA 组。
