High-throughput SuperSAGE for digital gene expression analysis of multiple samples using next generation sequencing.

PLoS One 2010 Aug 6;5(8):e12010. Epub 2010 Aug 6.

Iwate Biotechnology Research Center, Kitakami, Japan.

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.

Download full-text PDF

Source
http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0012010PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2917361PMC
August 2010
52 Reads
42 Citations
3.234 Impact Factor

Publication Analysis

Top Keywords

gene expression
12
digital gene
12
tags samples
4
discriminate tags
4
employed discriminate
4
sequences employed
4
examples provided
4
foreseen examples
4
allow researchers
4
applications foreseen
4
barcodes allow
4
samples barcodes
4
barcode sequences
4
protocol barcode
4
tag fragments
4
accurate digital
4
26-bp tag
4
isolation 26-bp
4
fragments expressed
4
study including
4

Similar Publications