Molecular haplotyping at high throughput
Molecular haplotyping at high throughput
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DOI：
10.1093/nar/gnf095
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发表时间：
2002-10-01
影响因子：
14.9
通讯作者：
Gut, IG
中科院分区：
文献类型：
--
作者：
Tost, J;Brandt, O;Gut, IG
单核苷酸多态性（SNP）的单倍型或等位基因相位的重建是旨在鉴定和解剖复杂遗传性状中涉及的遗传因素的研究的关键组成部分。在人类中，这通常涉及在病例/对照或其他队列中调查SNP，其中单倍型只能通过统计方法从基因型中部分推断，从而导致功率损失。此外，替代的统计方法可能导致对存在的最可能的单倍型的不同评估，以及当数据不明确时的不同单倍型频率估计。考虑到SNP研究的成本和复杂性，一种强大且易于使用的分子技术，允许直接从个体DNA样本中确定单倍型，将具有广泛的适用性。在这里，我们提出了一个可靠的，自动化的和高通量的方法，分子单倍型在2 kb，并可能更长的序列段，是基于物理测定的阶段的SNP等位基因的任何一个单独的父系单倍体。我们证明，分子单体型与这种技术是不复杂的SNP基因分型时，实施基质辅助激光解吸/电离质谱，我们还表明，该方法可以应用于其他DNA变异检测平台。分子单倍型是说明了充分描述的β（2）-肾上腺素能受体基因。
Reconstruction of haplotypes, or the allelic phase, of single nucleotide polymorphisms (SNPs) is a key component of studies aimed at the identification and dissection of genetic factors involved in complex genetic traits. In humans, this often involves investigation of SNPs in case/control or other cohorts in which the haplotypes can only be partially inferred from genotypes by statistical approaches with resulting loss of power. Moreover, alternative statistical methodologies can lead to different evaluations of the most probable haplotypes present, and different haplotype frequency estimates when data are ambiguous. Given the cost and complexity of SNP studies, a robust and easy-to-use molecular technique that allows haplotypes to be determined directly from individual DNA samples would have wide applicability. Here, we present a reliable, automated and high-throughput method for molecular haplotyping in 2 kb, and potentially longer, sequence segments that is based on the physical determination of the phase of SNP alleles on either of the individual paternal haploids. We demonstrate that molecular haplotyping with this technique is not more complicated than SNP genotyping when implemented by matrix-assisted laser desorption/ionisation mass spectrometry, and we also show that the method can be applied using other DNA variation detection platforms. Molecular haplotyping is illustrated on the well-described beta(2)-adrenergic receptor gene.