Abstract：
A major goal for biologists is to understand the connection between genes and phenotypic traits, and genetic mapping in experimental populations remains a powerful approach for discovering the causal genes underlying phenotypes. For genetic mapping, the process of genotyping was previously a major rate-limiting step. Modern sequencing technology has greatly improved the resolution and speed of genetic mapping by reducing the time, labor, and cost per genotyping marker. In addition, the ability to perform genotyping-by-sequencing (GBS) has facilitated large-scale population genetic analyses by providing a simpler way to survey segregating genetic variation in natural populations. Here we present two protocols for GBS, using the Illumina platform, that can be applied to a wide range of genotyping projects in different species. The first protocol is for genotyping a subset of marker positions genome-wide using restriction digestion, and the second is for preparing inexpensive paired-end whole-genome libraries. We discuss the suitability of each approach for different genotyping applications and provide notes for adapting these protocols for use with a liquid-handling robot.
摘要：
生物学家的主要目标是了解基因与表型性状之间的联系，并且在实验人群中进行遗传作图仍然是发现潜在表型的因果基因的有力方法。对于遗传作图，基因分型过程以前是一个主要的限速步骤.现代测序技术通过缩短时间,大大提高了遗传作图的分辨率和速度,劳动,和每个基因分型标记的成本。此外,通过提供一种更简单的方法来调查自然种群中的分离遗传变异，从而进行测序基因分型（GBS）的能力促进了大规模种群遗传分析。在这里，我们提出了两种GBS协议，使用Illumina平台，这可以应用于不同物种的广泛的基因分型项目。第一个方案是使用限制性消化在全基因组范围内对标记位置的子集进行基因分型，第二个是制备廉价的配对末端全基因组文库。我们讨论了每种方法对不同基因分型应用的适用性，并提供了适应这些协议以与液体处理机器人一起使用的注意事项。
