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HAPPY mapping was designed to pursue the analysis of approximately random HAPloid DNA breakage samples using the PolYmerase chain reaction for mapping genomes. In the present study, we improved the method and integrated two other molecular techniques into the process: whole genome amplification and the Sequenom SNP (single nucleotide polymorphism) genotyping assay in order to facilitate whole genome mapping of X. tropicalis. The former technique amplified enough DNA materials to genotype a large number of markers, while the latter allowed for relatively high throughput marker genotyping with multiplex assays on the HAPPY lines. A total of 58 X. tropicalis genes were genotyped on an initial panel of 383 HAPPY lines, which contributed to formation of a working panel of 146 lines. Further genotyping of 29 markers on the working panel led to construction of a HAPPY map for the X. tropicalis genome. We believe that our improved HAPPY method described in the present study has paved the way for the community to map different genomes with a simple, but powerful approach.
Figure 1. Effects of sampled fragment sizes on WGA yields (A) and marker retention rate (B). Data are presented as least square means ± standard error. The bars without common letters are significantly different (P<0.05).
Figure 2. Marker retention rate on whole genome DNA samples (A) and HAPPY lines (B).
Figure 3. UCE marker locations and orders on human chromosome 1 (Reference chromosome) as compared to their locations and orders on our frog HAPPY map (Dataset 2) and frog genome assembly v7.1 (Dataset 3).
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