small rna expression profiling by high-throughput sequencing implications of enzymatic manipulation小rna表达酶操纵由高通量测序分析的影响.pdf

Hindawi Publishing Corporation Journal of Nucleic Acids Volume 2012, Article ID 360358, 15 pages doi:10.1155/2012/360358 Review Article Small RNA Expression Profiling by High-Throughput Sequencing: Implications of Enzymatic Manipulation Fanglei Zhuang, Ryan T. Fuchs, and G. Brett Robb RNA Division, New England Biolabs Inc., 240 County Road, Ipswich, MA 01938, USA Correspondence should be addressed to G. Brett Robb, robb@ Received 2 December 2011; Accepted 5 March 2012 Academic Editor: Atsushi Maruyama Copyright © 2012 Fanglei Zhuang et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Eukaryotic regulatory small RNAs (sRNAs) play significant roles in many fundamental cellular processes. As such, they have emerged as useful biomarkers for diseases and cell differentiation states. sRNA-based biomarkers outperform traditional messenger RNA-based biomarkers by testing fewer targets with greater accuracy and providing earlier detection for disease states. Therefore, expression profiling of sRNAs is fundamentally important to further advance the understanding of biological processes, as well as diagnosis and treatment of diseases. High-throughput sequencing (HTS) is a powerful approach for both sRNA discovery and expression profiling. Here, we discuss the general considerations for sRNA-based HTS profiling methods from RNA preparation to sequencing library construction, with a focus on the causes of systematic error. By examining the enzymatic manipulation steps of sRNA expression profiling, this paper aims to demystify current HTS-based sRNA profiling approaches and to aid researchers in the inf