The efficacy of Cot-based gene enrichment in wheat (Triticum aestivum L.). |
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Authors: | Didier Lamoureux Daniel G Peterson Wanlong Li John P Fellers Bikram S Gill |
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Affiliation: | Wheat Genetics Resource Center, Department of Plant Pathology, Kansas State University, Manhattan 66506, USA. |
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Abstract: | We report the results of a study on the effectiveness of Cot filtration (CF) in the characterization of the gene space of bread wheat (Triticum aestivum L.), a large genome species (1C = 16,700 Mb) of tremendous agronomic importance. Using published Cot data as a guide, 2 genomic libraries for hexaploid wheat were constructed from the single-stranded DNA collected at Cot values > 1188 and 1639 M x s. Compared with sequences from a whole genome shotgun library from Aegilops tauschii (the D genome donor of bread wheat), the CF libraries exhibited 13.7-fold enrichment in genes, 5.8-fold enrichment in unknown low-copy sequences, and a 3-fold reduction in repetitive DNA. CF is twice as efficient as methylation filtration at enriching wheat genes. This research suggests that, with improvements, CF will be a highly useful tool in sequencing the gene space of wheat. |
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