Journal article
High-throughput SuperSAGE for digital gene expression analysis of multiple samples using next generation sequencing
PLoS ONE, Vol.5(8), e12010
2010
Abstract
We established a protocol of the SuperSAGE technology combined with next-generation sequencing, coined "High- Throughput (HT-) SuperSAGE". SuperSAGE is a method of digital gene expression profiling that allows isolation of 26-bp tag fragments from expressed transcripts. In the present protocol, index (barcode) sequences are employed to discriminate tags from different samples. Such barcodes allow researchers to analyze digital tags from transcriptomes of many samples in a single sequencing run by simply pooling the libraries. Here, we demonstrated that HT-SuperSAGE provided highly sensitive, reproducible and accurate digital gene expression data. By increasing throughput for analysis in HT-SuperSAGE, various applications are foreseen and several examples are provided in the present study, including analyses of laser-microdissected cells, biological replicates and tag extraction using different anchoring enzymes.
Details
- Title
- High-throughput SuperSAGE for digital gene expression analysis of multiple samples using next generation sequencing
- Authors/Creators
- H. Matsumura (Author/Creator) - Shinshu UniversityK. Yoshida (Author/Creator) - Iwate Biotechnology Research CenterS. Luo (Author/Creator) - Illumina (United States)E. Kimura (Author/Creator) - Iwate Medical UniversityT. Fujibe (Author/Creator) - Iwate Biotechnology Research CenterZ. Albertyn (Author/Creator) - Murdoch UniversityR.A. Barrero (Author/Creator) - Murdoch UniversityD.H. Krüger (Author/Creator) - Charité - Universitätsmedizin BerlinG. Kahl (Author/Creator) - Goethe University FrankfurtG.P. Schroth (Author/Creator) - Illumina (United States)R. Terauchi (Author/Creator) - Iwate Biotechnology Research Center
- Publication Details
- PLoS ONE, Vol.5(8), e12010
- Identifiers
- 991005540396707891
- Murdoch Affiliation
- Centre for Comparative Genomics
- Language
- English
- Resource Type
- Journal article
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