Strategies and Best Practice in Cloning Small RNAs.
Strategies and Best Practice in Cloning Small RNAs.
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发表时间：
2020-08-03
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通讯作者：
Gu W
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作者：
Dai H;Gu W
高通量测序因其灵敏性和便捷性已成为分析核酸的标准和有力工具。小rna在调节细胞和病毒基因方面发挥着重要作用。传统的小RNA分析方法繁琐，对许多小RNA种类往往缺乏准确性、特异性和敏感性。因此，高通量测序成为分析小rna不可缺少的工具。然而，为高通量测序生成可靠且具有代表性的小RNA文库是具有挑战性的，因为小RNA通常表达水平极低，并且通常包含影响文库构建的修饰，通常会导致偏读。本文比较了制备高质量、高可靠性小RNA文库的各种策略，并对制备高通量测序RNA文库的最佳实践提出了建议。
High-throughput sequencing has become a standard and powerful tool for analyzing nucleic acids primarily due to its sensitivity and convenience. Small RNAs play important roles in regulating cellular and viral genes. The conventional methods for small RNA analyses are tedious and often lack accuracy, specificity and sensitivity for many small RNA species. Therefore, high-throughput sequencing becomes an indispensable tool for analyzing small RNAs. However, it is challenging to generate a reliable and representative small RNA library for high-throughput sequencing since small RNAs are usually expressed at extremely low levels and often contain modifications which affect library construction, usually causing biased readouts. This review compares various strategies for generating small RNA libraries of high quality and reliability, and provides recommendations on best practice in preparing high-throughput sequencing RNA libraries.