Department of Agronomy, Iowa State University, Ames, Iowa, United States of America.
PLoS One. 2010 Dec 2;5(12):e14178. doi: 10.1371/journal.pone.0014178.
To date, microarray-based genotyping of large, complex plant genomes has been complicated by the need to perform genome complexity reduction to obtain sufficiently strong hybridization signals. Genome complexity reduction techniques are, however, tedious and can introduce unwanted variables into genotyping assays. Here, we report a microarray-based genotyping technology for complex genomes (such as the 2.3 GB maize genome) that does not require genome complexity reduction prior to hybridization. Approximately 200,000 long oligonucleotide probes were identified as being polymorphic between the inbred parents of a mapping population and used to genotype two recombinant inbred lines. While multiple hybridization replicates provided ∼97% accuracy, even a single replicate provided ∼95% accuracy. Genotyping accuracy was further increased to >99% by utilizing information from adjacent probes. This microarray-based method provides a simple, high-density genotyping approach for large, complex genomes.
迄今为止,基于微阵列的大型复杂植物基因组基因分型一直受到需要进行基因组复杂度降低以获得足够强的杂交信号的限制。然而,基因组复杂度降低技术繁琐,并且可能会给基因分型实验引入不必要的变量。在这里,我们报告了一种用于复杂基因组(如 23 亿碱基对的玉米基因组)的基于微阵列的基因分型技术,该技术在杂交之前不需要进行基因组复杂度降低。大约 200000 个长寡核苷酸探针被鉴定为在一个作图群体的自交亲本之间具有多态性,并用于对两个重组自交系进行基因分型。虽然多个杂交重复提供了约 97%的准确性,但即使单个重复也提供了约 95%的准确性。通过利用相邻探针的信息,基因分型的准确性进一步提高到>99%。这种基于微阵列的方法为大型复杂基因组提供了一种简单、高密度的基因分型方法。