title: The (in)complete organelle genome: exploring the use and nonuse of available technologies for characterizing mitochondrial and plastid chromosomes.
abstract: Not long ago, scientists paid dearly in time, money and skill for every nucleotide that they sequenced. Today, DNA sequencing technologies epitomize the slogan 'faster, easier, cheaper and more', and in many ways, sequencing an entire genome has become routine, even for the smallest laboratory groups. This is especially true for mitochondrial and plastid genomes. Given their relatively small sizes and high copy numbers per cell, organelle DNAs are currently among the most highly sequenced kind of chromosome. But accurately characterizing an organelle genome and the information it encodes can require much more than DNA sequencing and bioinformatics analyses. Organelle genomes can be surprisingly complex and can exhibit convoluted and unconventional modes of gene expression. Unravelling this complexity can demand a wide assortment of experiments, from pulsed-field gel electrophoresis to Southern and Northern blots to RNA analyses. Here, we show that it is exactly these types of 'complementary' analyses that are often lacking from contemporary organelle genome papers, particularly short 'genome announcement' articles. Consequently, crucial and interesting features of organelle chromosomes are going undescribed, which could ultimately lead to a poor understanding and even a misrepresentation of these genomes and the genes they express. High-throughput sequencing and bioinformatics have made it easy to sequence and assemble entire chromosomes, but they should not be used as a substitute for or at the expense of other types of genomic characterization methods.
authors.affiliation: Department of Biology, University of Western Ontario, London, Ontario, Canada, N6A 5B7.
authors.forename: Matheus
authors.initials: M
authors.lastname: Sanitá Lima
authors.affiliation: Department of Biology, University of Western Ontario, London, Ontario, Canada, N6A 5B7.
authors.forename: Laura C
authors.initials: LC
authors.lastname: Woods
authors.affiliation: Department of Biology, University of Western Ontario, London, Ontario, Canada, N6A 5B7.
authors.forename: Matthew W
authors.initials: MW
authors.lastname: Cartwright
authors.affiliation: Department of Biology, University of Western Ontario, London, Ontario, Canada, N6A 5B7. dsmit242@uwo.ca.
authors.forename: David Roy
authors.initials: DR
authors.lastname: Smith
mesh_terms: D019295:Computational Biology / Q000379:methods; D023281:Genomics / Q000379:methods*; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D008928:Mitochondria / Q000235:genetics*; D018087:Plastids / Q000235:genetics*; D017422:Sequence Analysis, DNA / Q000379:methods*
keywords: RNA-Seq; genome announcement; genome report; next-generation sequencing; transcriptomics