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通过纳米孔测序确定复杂mRNA中的外显子连接性。

Determining exon connectivity in complex mRNAs by nanopore sequencing.

作者信息

Bolisetty Mohan T, Rajadinakaran Gopinath, Graveley Brenton R

机构信息

Department of Genetics and Genome Sciences, Institute for Systems Genomics, University of Connecticut Health Center, Farmington, CT, 06030, USA.

Present Address: The Jackson Laboratory for Genomic Medicine, Farmington, CT, 06030, USA.

出版信息

Genome Biol. 2015 Sep 30;16:204. doi: 10.1186/s13059-015-0777-z.

Abstract

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.

摘要

短读长高通量RNA测序虽然功能强大,但在直接测量包含多个距离超过最大读长的可变外显子的mRNA中的外显子连接性方面能力有限。在这里,我们使用牛津纳米孔MinION测序仪来鉴定从四个果蝇基因Dscam1、MRP、Mhc和Rdl表达的7899种“全长”异构体。这些结果表明,纳米孔测序可用于解析单个异构体,并且它有可能成为全面转录组表征的强大方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/46ed/4588896/840c368289c2/13059_2015_777_Fig1_HTML.jpg

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