Digital gene expression by tag sequencing on the illumina genome analyzer.
This unit provides a protocol for performing digital gene expression profiling on the Illumina Genome Analyzer sequencing platform. Tag sequencing (Tag-seq) is an implementation of the LongSAGE protocol on the Illumina sequencing platform that increases utility while reducing both the cost and time required to generate gene expression profiles. The ultra-high-throughput sequencing capability of the Illumina platform allows the cost-effective generation of libraries containing an average of 20 million tags, a 200-fold improvement over classical LongSAGE. Tag-seq has less sequence composition bias, leading to a better representation of AT-rich tag sequences, and allows a more accurate profiling of a subset of the transcriptome characterized by AT-rich genes expressed at levels below the threshold of detection of LongSAGE (Morrissy et al., 2009).
Morrissy
Sorana
S
BCCA Genome Sciences Centre, University of British Columbia, Vancouver, British Columbia, Canada.
Zhao
Yongjun
Y
Delaney
Allen
A
Asano
Jennifer
J
Dhalla
Noreen
N
Li
Irene
I
McDonald
Helen
H
Pandoh
Pawan
P
Prabhu
Anna-Liisa
AL
Tam
Angela
A
Hirst
Martin
M
Marra
Marco
M
D020224:Expressed Sequence Tags; D020869:Gene Expression Profiling / Q000379:methods*; D015723:Gene Library; D023281:Genomics / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods; D012333:RNA, Messenger / Q000235:genetics*; D017422:Sequence Analysis, DNA / Q000379:methods*
D012333:RNA, Messenger