Seeb James E, Pascal Carita E, Ramakrishnan Ramesh, Seeb Lisa W
School of Aquatic and Fishery Sciences, University of Washington, Seattle, WA, USA.
Methods Mol Biol. 2009;578:277-92. doi: 10.1007/978-1-60327-411-1_18.
Population genetics studies play an increasingly important role in the management and conservation of nonmodel organisms. Unlike studies with model organisms, a typical population genetics study of a nonmodel organism may be conducted by analyzing thousands or hundreds of thousands of individuals for several dozen single nucleotide polymorphisms (SNPs). The use of robust, robotically mediated TaqMan reactions provides substantial advantages in these types of studies. We describe the methods and laboratory setup for analyzing a sustained high throughput of SNP assays in routine university or natural resource agency laboratories with a handful of thermal cyclers. Agencies sustain rates of nearly 150,000 assays per week using uniplex reactions with the Applied Biosystems 7900HT Fast Real-Time PCR System (AB 7900HT). We further describe the medium-density array run on the BioMark from Fluidigm, which increases this rate to over 500,000 assays per week by multiplexing 96 samples for 96 SNPs.
群体遗传学研究在非模式生物的管理和保护中发挥着越来越重要的作用。与模式生物的研究不同，对非模式生物进行典型的群体遗传学研究可能需要分析数千或数十万个体的几十种单核苷酸多态性（SNP）。在这类研究中，使用强大的、由机器人介导的TaqMan反应具有显著优势。我们描述了在配备少量热循环仪的常规大学或自然资源机构实验室中分析SNP检测持续高通量的方法和实验室设置。各机构使用应用生物系统公司的7900HT快速实时PCR系统（AB 7900HT）进行单重反应，每周可进行近150,000次检测。我们还描述了在Fluidigm公司的BioMark上运行的中密度阵列，通过对96个样本的96个SNP进行多重分析，将这一速率提高到每周超过500,000次检测。
