Keam Simon P, Sobala Andrew, Humphreys David T, Suter Catherine M, Hutvagner Gyorgy
Faculty of Engineering and Information Technology, Centre of Health Technologies, University of Technology, Sydney, 235 Jones Street, Ultimo, Sydney, NSW, 2007, Australia.
Methods Mol Biol. 2014;1173:157-67. doi: 10.1007/978-1-4939-0931-5_14.
The rapidly growing list of small RNA species generated by next-generation sequencing technologies has accelerated the development of new bioinformatics tools for their detection. Small RNAs generated from tRNAs, transfer RNA-derived fragments (tRFs), represent a novel challenge in accurately identifying and distinguishing them from random degradation products of tRNAs. Here, we describe a bioinformatics approach to detect tRFs in next-generation sequencing libraries. We also present a biochemical purification protocol for enriching 5' tRFs and separating them from miRNAs. And finally, we suggest reliable methods for detecting and quantifying tRFs.
下一代测序技术产生的小RNA种类迅速增加，加速了用于检测它们的新生物信息学工具的开发。从tRNA产生的小RNA，即转移RNA衍生片段（tRF），在准确识别它们并将其与tRNA的随机降解产物区分开来方面提出了新的挑战。在这里，我们描述了一种在下一代测序文库中检测tRF的生物信息学方法。我们还提出了一种生化纯化方案，用于富集5' tRF并将它们与miRNA分离。最后，我们提出了检测和定量tRF的可靠方法。
