Matsumura Hideo, Krüger Detlev H, Kahl Günter, Terauchi Ryohei
Gene Research Center, Shinshu University, 3-15-1 Tokita, Ueda, Nagano, 386-8567, Japan,
Methods Mol Biol. 2015;1236:181-95. doi: 10.1007/978-1-4939-1743-3_14.
SuperSAGE is a tag-based transcript profiling method, which allows to analyze the expression of thousands of genes at a time. In SuperSAGE, 26 bp tags are extracted from cDNA using the type III restriction enzyme, EcoP15I. In SuperSAGE, the amount of transcripts was represented by tag counts. Taking advantage of uniqueness of the 26 bp tags, host and virus transcripts can be monitored in virus-infected cells. Combining next generation sequencing technology, we established High-throughput SuperSAGE (Ht-SuperSAGE), which allows the analysis of multiple samples with reduced time and cost. In this chapter, we present the protocol of Ht-SuperSAGE involving a recently available benchtop type next generation sequencer.
超级SAGE是一种基于标签的转录本分析方法,它能够一次性分析数千个基因的表达情况。在超级SAGE中,使用III型限制性内切酶EcoP15I从cDNA中提取26bp的标签。在超级SAGE中,转录本的数量由标签计数表示。利用26bp标签的独特性,可以在病毒感染的细胞中监测宿主和病毒转录本。结合下一代测序技术,我们建立了高通量超级SAGE(Ht-SuperSAGE),它能够以更少的时间和成本分析多个样本。在本章中,我们介绍了涉及最近可用的台式下一代测序仪的Ht-SuperSAGE方案。