Plant Genotyping pp 169-182

Part of the Methods in Molecular Biology book series (MIMB, volume 1245) | Cite as

Genomic Reduction Assisted Single Nucleotide Polymorphism Discovery Using 454-Pyrosequencing

  • Peter J. Maughan
  • Joshua A. Udall
  • Eric N. Jellen
Protocol

Abstract

We report the development of a simple genomic reduction protocol based on 454-pyrosequencing technology that discovers large numbers of single nucleotide polymorphisms (SNP) from pooled DNA samples. The method is based on the conservation of restriction endonuclease sites across samples and biotin separation for genomic reduction and the addition of multiplex identifier (MID) barcodes to each of the pooled samples to allow for postsequencing deconvolution of the pooled DNA fragments and SNP discovery.

Key words

Single nucleotide polymorphism (SNP) 454-Pyrosequencing Genomic reduction 

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Peter J. Maughan
    • 1
  • Joshua A. Udall
    • 2
  • Eric N. Jellen
    • 3
  1. 1.5144 LSB, Department of Plant and Wildlife SciencesBrigham Young UniversityProvoUSA
  2. 2.5133 LSB, Department of Plant and Wildlife SciencesBrigham Young UniversityProvoUSA
  3. 3.4105B LSB, Department of Plant and Wildlife SciencesBrigham Young UniversityProvoUSA

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