title: Identification of factors required for meristem function in Arabidopsis using a novel next generation sequencing fast forward genetics approach.
abstract: BACKGROUND
  Phenotype-driven forward genetic experiments are powerful approaches for linking phenotypes to genomic elements but they still involve a laborious positional cloning process. Although sequencing of complete genomes now becomes available, discriminating causal mutations from the enormous amounts of background variation remains a major challenge.
  
  
  METHODS
  To improve this, we developed a universal two-step approach, named 'fast forward genetics', which combines traditional bulk segregant techniques with targeted genomic enrichment and next-generation sequencing technology
  
  
  RESULTS
  As a proof of principle we successfully applied this approach to two Arabidopsis mutants and identified a novel factor required for stem cell activity.
  
  
  CONCLUSIONS
  We demonstrated that the 'fast forward genetics' procedure efficiently identifies a small number of testable candidate mutations. As the approach is independent of genome size, it can be applied to any model system of interest. Furthermore, we show that experiments can be multiplexed and easily scaled for the identification of multiple individual mutants in a single sequencing run.
authors: Hubrecht Institute, Developmental Biology and Stem Cell Research, KNAW and University Medical Center Utrecht, Uppsalalaan 8, Utrecht, The Netherlands.
authors: Michal
authors: M
authors: Mokry
authors: Isaäc J
authors: IJ
authors: Nijman
authors: Anja
authors: A
authors: van Dijken
authors: Rene
authors: R
authors: Benjamins
authors: Renze
authors: R
authors: Heidstra
authors: Ben
authors: B
authors: Scheres
authors: Edwin
authors: E
authors: Cuppen
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