Sysmex Partec 倍性分析仪在植物育种上的应用---倍体分析仪测定植物细胞核DNA含量
Abstract：
Flow cytometry (FCM) using DNA-selective fluorochromes is now the prevailing method for the measurement of nuclear DNA content in plants. Ease of sample preparation and high sample throughput make it generally better suited than other methods such as Feulgen densitometry to estimate genome size, level of generative polyploidy, nuclear replication state and endopolyploidy (polysomaty). Here we present four protocols for sample preparation (suspensions of intact cell nuclei) and describe the analysis of nuclear DNA amounts using FCM. We consider the chemicals and equipment necessary, the measurement process, data analysis, and describe the most frequent problems encountered with plant material such as the interference of secondary metabolites. The purpose and requirement of internal and external standardization are discussed. The importance of using a correct terminology for DNA amounts and genome size is underlined, and its basic principles are explained.
摘要：
使用DNA选择性荧光染料的倍体分析仪（FCM）现在是测量植物DNA含量的主流方法。样品制备的简便性和高样品通量使其通常比其他方法（如福尔根密度测定法）更适合估计基因组大小、生殖多倍体水平、核复制状态和内多倍性（多聚体）。在这里，我们提出了四种样品制备方案（完整细胞核的悬浮液），并描述了使用FCM分析核DNA量。我们考虑了必要的化学品和设备、测量过程、数据分析，并描述了植物材料遇到的最常见问题，如次生代谢物的干扰。讨论了内外部标准化的目的和要求。强调了使用正确的术语来描述DNA数量和基因组大小的重要性，并解释了其基本原理。
关键词：
Sysmex Partec 倍性分析仪，多倍体诱导仪，静水压仪，CyFlow Ploidy Analyser 流式细胞仪，DNA含量测定
