Combining DNA pooling with selective recombinant genotyping for increased efficiency in fine mapping.
One of the key steps in positional cloning and marker-aided selection is to identify marker(s) tightly linked to the target gene (i.e., fine mapping). Selective genotyping such as selective recombinant genotyping (SRG) is commonly used in fine mapping for cost-saving. To further decrease genotyping effort and rapidly screen for tightly linked markers, we propose here a combined DNA pooling and SRG strategy. A two-stage pooled genotyping can be used for identifying recombinants between a pair of flanking markers more efficiently, and a joint use of bulked DNA analysis and two-stage pooling can also save cost for genotyping recombinants. The combined DNA pooling and SRG strategy can further be extended to fine mapping for polygenic traits. The numerical results based on hypothetical scenarios and an illustrative application to fine mapping of a mutant gene, called xl(t), in rice suggest that the proposed strategy can remarkably reduce genotyping amount compared with the conventional SRG.
Chi
Xiao-Fei
XF
Institute of Nuclear Agricultural Sciences, Zhejiang University, Hangzhou, People's Republic of China.
Lou
Xiang-Yang
XY
Shu
Qing-Yao
QY
D002874:Chromosome Mapping / Q000379:methods*; D018744:DNA, Plant / Q000235:genetics; D005787:Gene Frequency; D005838:Genotype; D012275:Oryza / Q000235:genetics*; D040641:Quantitative Trait Loci
D018744:DNA, Plant