Genomic Reduction Assisted Single Nucleotide Polymorphism Discovery Using 454-Pyrosequencing
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 wordsSingle nucleotide polymorphism (SNP) 454-Pyrosequencing Genomic reduction
This research was funded by the Ezra Taft Benson Agriculture and Food Institute. We gratefully acknowledge Dr. Edward Wilcox (BYU) for his assistance and advice with regards to 454-pyrosequencing.
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