Transcriptome profiling using single-molecule direct RNA sequencing.
作者： Fatih Ozsolak , Patrice M. Milos
DOI: 10.1007/978-1-61779-089-8_4
关键词:
摘要: Methods for in-depth characterization of transcriptomes and quantification transcript levels have emerged as valuable tools understanding cellular physiology human disease biology, begun to be utilized in various clinical diagnostic applications. Today, current methods by the scientific community typically require RNA converted cDNA prior comprehensive measurements. However, this conversion process has been shown introduce many biases artifacts that interfere with proper quantitation transcripts. We developed a direct sequencing (DRS) approach, which, unlike other technologies, is sequenced directly without cDNA. The benefits DRS include ability use minute quantities (e.g. on order several femtomoles) minimal sample preparation, analyze short RNAs which pose unique challenges analysis using cDNA-based approaches, perform these analyses low-cost high-throughput manner. Here, we describe strategies procedures employ prepare species DRS.
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