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Methods for obtaining and analyzing whole chloroplast genome sequences

  • Author(s): Jansen, Robert K.
  • Raubeson, Linda A.
  • Boore, Jeffrey L.
  • dePamphilis, Claude W.
  • Chumley, Timothy W.
  • Haberle, Romemarie C.
  • Wyman, Stacia K.
  • Alverson, Andrew J.
  • Peery, Rhiannon
  • Herman, Sallie J.
  • Fourcade, H. Matthew
  • Kuehl, Jennifer V.
  • McNeal, Joel R.
  • Leebens-Mack, James
  • Cui, Liying
  • et al.
Abstract

During the past decade there has been a rapid increase in our understanding of plastid genome organization and evolution due to the availability of many new completely sequenced genomes. Currently there are 43 complete genomes published and ongoing projects are likely to increase this sampling to nearly 200 genomes during the next five years. Several groups of researchers including ours have been developing new techniques for gathering and analyzing entire plastid genome sequences and details of these developments are summarized in this chapter. The most important recent developments that enhance our ability to generate whole chloroplast genome sequences involve the generation of pure fractions of chloroplast genomes by whole genome amplification using rolling circular amplification, cloning genomes into Fosmid or BAC vectors, and the development of an organellar annotation program (DOGMA). In addition to providing details of these methods, we provide an overview of methods for analyzing complete plastid genome sequences for repeats and gene content, as well as approaches for using gene order and sequence data for phylogeny reconstruction. This explosive increase in the number of sequenced plastid genomes and improved computational tools will provide many insights into the evolution of these genomes and much new data for assessing relationships at deep nodes in plants and other photosynthetic organisms.

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