Hardie David C, Gregory T Ryan, Hebert Paul D N
Department of Zoology, University of Guelph, Guelph, Ontario, Canada.
J Histochem Cytochem. 2002 Jun;50(6):735-49. doi: 10.1177/002215540205000601.
The study of genome size variation is important from a number of practical and theoretical perspectives. For example, the long-standing "C-value enigma" relating to the more than 200,000-fold range in eukaryotic genome sizes is best studied from a broad comparative standpoint. Genome size data are also required in detailed analyses of genome structure and evolution. The choice of future genome sequencing projects will be dependent on knowledge regarding the sizes of genomes to be sequenced, and so on. To date, genome size data have been acquired primarily by Feulgen microdensitometry or flow cytometry. Each has several advantages but also important limitations. In this review, we provide a practical guide to the new technique of Feulgen image analysis densitometry. The review is designed for those interested in genome size measurements but not extensively experienced in histochemistry, densitometry, or microscopy. Therefore, relevant historical and technical background information is included. For easy reference, we provide recipes for required reagents, guidelines for cell staining, and a checklist of steps for successful image analysis. We hope that the accuracy, rapidity, and cost-effectiveness of Feulgen image analysis demonstrated here will stimulate further surveys of genome sizes in a variety of taxa.
从多个实践和理论角度来看，基因组大小变异的研究都很重要。例如，与真核生物基因组大小超过20万倍的范围相关的长期存在的“C值谜”，最好从广泛的比较角度进行研究。在基因组结构和进化的详细分析中也需要基因组大小数据。未来基因组测序项目的选择将取决于有关待测序基因组大小等方面的知识。迄今为止，基因组大小数据主要通过福尔根显微密度测定法或流式细胞术获得。每种方法都有几个优点，但也有重要的局限性。在本综述中，我们为福尔根图像分析密度测定法这项新技术提供了实用指南。该综述是为那些对基因组大小测量感兴趣但在组织化学、密度测定法或显微镜方面经验不多的人设计的。因此，包含了相关的历史和技术背景信息。为方便参考，我们提供了所需试剂的配方、细胞染色指南以及成功进行图像分析的步骤清单。我们希望这里展示的福尔根图像分析的准确性、快速性和成本效益将激发对各种分类群基因组大小的进一步研究。
