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Mol Ecol Resour. 2013 Jul;13(4):551-8. doi: 10.1111/1755-0998.12107. Epub 2013 Apr 25.

Of contigs and quagmires: next-generation sequencing pitfalls associated with transcriptomic studies.

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  • 1Department of Biological Sciences, Purdue University, West Lafayette, IN 47907, USA. dewoody@purdue.edu

Abstract

Molecular ecologists have good reasons to be excited about the newest DNA/RNA sequencing technologies. However, this exuberance should be tempered with a hefty dose of reality: new sequencing technologies come with significant new challenges. Herein, we offer a brief overview of some practical problems encountered during transcriptomics studies conducted in our laboratory, and of nontrivial issues that prospective practitioners should consider. These include template contamination (e.g. from xenobiotics) and the cutting-room floor problem, whereby most of the data are often unassembled, unannotated and unused. We also highlight computational requirements, including hardware, personnel time and associated skill sets. We are very optimistic about the future of molecular ecology, but we hope this cautionary overview will help neophytes better recognize some key challenges associated with new technologies.

© 2013 John Wiley & Sons Ltd.

[PubMed - indexed for MEDLINE]
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