Preparation of Small RNA NGS Libraries from Biofluids
Preparation of Small RNA NGS Libraries from Biofluids
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DOI：
10.1007/978-1-4939-7652-2_13
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发表时间：
2018-01-01
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影响因子：
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通讯作者：
Galas, David
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文献类型：
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作者：
Etheridge, Alton;Wang, Kai;Galas, David
下一代测序（NGS）是一种强大的转录组分析方法。与其他基因表达谱分析方法（例如微阵列）不同，NGS 提供了额外的信息，例如剪接变体、序列多态性和新转录本。因此，NGS 非常适合对生物体液中各种细胞外 RNA (exRNA) 进行全面分析。 ExRNA 因其在细胞间通讯中可能发挥的生物学作用以及作为生物标志物或用于治疗目的的潜在用途而引起了极大的兴趣。在这里，我们描述了用于 NGS 分析的小 RNA 文库的修改方案。该方案已经过优化，可用于含有低输入 exRNA 的样品，例如血浆或血清，并进行了修改，旨在减少商业小 RNA 文库构建试剂盒通常遇到的序列特异性偏差。
Next generation sequencing (NGS) is a powerful method for transcriptome analysis. Unlike other gene expression profiling methods, such as microarrays, NGS provides additional information such as splicing variants, sequence polymorphisms, and novel transcripts. For this reason, NGS is well suited for comprehensive profiling of the wide range of extracellular RNAs (exRNAs) in biofluids. ExRNAs are of great interest because of their possible biological role in cell-to-cell communication and for their potential use as biomarkers or for therapeutic purposes. Here, we describe a modified protocol for preparation of small RNA libraries for NGS analysis. This protocol has been optimized for use with low-input exRNA-containing samples, such as plasma or serum, and has modifications designed to reduce the sequence-specific bias typically encountered with commercial small RNA library construction kits.