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Author:
Date:
2014-11-05
[Abstract] High-throughput sequencing is a powerful tool for exploring small RNA populations in plants. The ever-increasing output from an Illumina Sequencing System allows for multiplexing multiple samples while still obtaining sufficient data for small RNA discovery and characterization. Here we describe a protocol for generating multiplexed small RNA libraries for sequencing up to 12 samples in one lane of an Illumina HiSeq System single-end, 50 base pair run. RNA ligases are used to add the 3’ and 5’ adaptors to purified small RNAs; ligation products that lack a small RNA molecule (adaptor-adaptor ...
[摘要] 高通量测序是探索植物中小RNA群体的强大工具。 来自Illumina测序系统的不断增加的输出允许多重多个样品，同时仍然获得足够的数据用于小RNA发现和表征。 在这里我们描述了一个协议，用于生成多重小RNA文库，用于在Illumina HiSeq系统单端，50碱基对运行的一个泳道中测序多达12个样品。 RNA连接酶用于将3'和5'接头添加至纯化的小RNA; 缺少小RNA分子（衔接子 - 衔接子产物）的连接产物被故意耗尽。 cDNA合成后，线性PCR步骤扩增DNA片段。 本文使用的3'PCR引物包括独特的6-核苷酸序列，以允许多达多达12个样品。
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