Title |
Stem cell transcriptome profiling via massive-scale mRNA sequencing
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Published in |
Nature Methods, May 2008
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DOI | 10.1038/nmeth.1223 |
Pubmed ID | |
Authors |
Nicole Cloonan, Alistair R R Forrest, Gabriel Kolle, Brooke B A Gardiner, Geoffrey J Faulkner, Mellissa K Brown, Darrin F Taylor, Anita L Steptoe, Shivangi Wani, Graeme Bethel, Alan J Robertson, Andrew C Perkins, Stephen J Bruce, Clarence C Lee, Swati S Ranade, Heather E Peckham, Jonathan M Manning, Kevin J McKernan, Sean M Grimmond |
Abstract |
We developed a massive-scale RNA sequencing protocol, short quantitative random RNA libraries or SQRL, to survey the complexity, dynamics and sequence content of transcriptomes in a near-complete fashion. This method generates directional, random-primed, linear cDNA libraries that are optimized for next-generation short-tag sequencing. We surveyed the poly(A)(+) transcriptomes of undifferentiated mouse embryonic stem cells (ESCs) and embryoid bodies (EBs) at an unprecedented depth (10 Gb), using the Applied Biosystems SOLiD technology. These libraries capture the genomic landscape of expression, state-specific expression, single-nucleotide polymorphisms (SNPs), the transcriptional activity of repeat elements, and both known and new alternative splicing events. We investigated the impact of transcriptional complexity on current models of key signaling pathways controlling ESC pluripotency and differentiation, highlighting how SQRL can be used to characterize transcriptome content and dynamics in a quantitative and reproducible manner, and suggesting that our understanding of transcriptional complexity is far from complete. |
X Demographics
Geographical breakdown
Country | Count | As % |
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Australia | 3 | 38% |
United States | 3 | 38% |
Unknown | 2 | 25% |
Demographic breakdown
Type | Count | As % |
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Scientists | 5 | 63% |
Members of the public | 3 | 38% |
Mendeley readers
Geographical breakdown
Country | Count | As % |
---|---|---|
United States | 44 | 3% |
United Kingdom | 20 | 1% |
Germany | 15 | 1% |
France | 8 | <1% |
Brazil | 8 | <1% |
Italy | 7 | <1% |
Sweden | 5 | <1% |
Norway | 3 | <1% |
Spain | 3 | <1% |
Other | 29 | 2% |
Unknown | 1297 | 90% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Researcher | 394 | 27% |
Student > Ph. D. Student | 342 | 24% |
Student > Master | 156 | 11% |
Professor > Associate Professor | 100 | 7% |
Student > Bachelor | 71 | 5% |
Other | 237 | 16% |
Unknown | 139 | 10% |
Readers by discipline | Count | As % |
---|---|---|
Agricultural and Biological Sciences | 777 | 54% |
Biochemistry, Genetics and Molecular Biology | 237 | 16% |
Medicine and Dentistry | 92 | 6% |
Computer Science | 42 | 3% |
Engineering | 27 | 2% |
Other | 97 | 7% |
Unknown | 167 | 12% |