Determining exon connectivity in complex mRNAs by nanopore sequencing.
Determining exon connectivity in complex mRNAs by nanopore sequencing.
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DOI：
10.1186/s13059-015-0777-z
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发表时间：
2015-09-30
期刊：
影响因子：
12.3
通讯作者：
Graveley BR
中科院分区：
文献类型：
--
作者：
Bolisetty MT;Rajadinakaran G;Graveley BR
短读长高通量 RNA 测序虽然功能强大，但其直接测量 mRNA 中外显子连接性的能力受到限制，这些 mRNA 包含多个距离远于最大读长的替代外显子。在这里，我们使用 Oxford Nanopore MinION 测序仪鉴定了 4 个果蝇基因 Dscam1、MRP、Mhc 和 Rdl 表达的 7,899 个“全长”亚型。这些结果表明，纳米孔测序可用于解卷积单个异构体，并且有潜力成为全面转录组表征的强大方法。
Short-read high-throughput RNA sequencing, though powerful, is limited in its ability to directly measure exon connectivity in mRNAs that contain multiple alternative exons located farther apart than the maximum read length. Here, we use the Oxford Nanopore MinION sequencer to identify 7,899 ‘full-length’ isoforms expressed from four Drosophila genes, Dscam1, MRP, Mhc, and Rdl. These results demonstrate that nanopore sequencing can be used to deconvolute individual isoforms and that it has the potential to be a powerful method for comprehensive transcriptome characterization.