Highly parallel direct RNA sequencing on an array of nanopores
Highly parallel direct RNA sequencing on an array of nanopores
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DOI：
10.1038/nmeth.4577
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发表时间：
2018-03-01
期刊：
影响因子：
48
通讯作者：
Turner, Daniel J.
中科院分区：
文献类型：
--
作者：
Garalde, Daniel R.;Snell, Elizabeth A.;Turner, Daniel J.
对生物样品中的RNA进行测序可以解锁大量信息，包括细菌和病毒的身份，选择性剪接的细微差别或生物体的转录状态。然而，目前的方法由于短读段长度和逆转录或扩增偏差而具有局限性。在这里，我们展示了纳米孔直接RNA-seq，这是一种高度平行，实时，单分子方法，可以绕过逆转录或扩增步骤。该方法产生全长、链特异性RNA序列，并能够直接检测RNA中的核苷酸类似物。
Sequencing the RNA in a biological sample can unlock a wealth of information, including the identity of bacteria and viruses, the nuances of alternative splicing or the transcriptional state of organisms. However, current methods have limitations due to short read lengths and reverse transcription or amplification biases. Here we demonstrate nanopore direct RNA-seq, a highly parallel, real-time, single-molecule method that circumvents reverse transcription or amplification steps. This method yields full-length, strand-specific RNA sequences and enables the direct detection of nucleotide analogs in RNA.