Sysmex Partec 倍性分析儀在植物育種上的應(yīng)用---倍體分析儀測(cè)定植物細(xì)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選擇性熒光染料的倍體分析儀（FCM）現(xiàn)在是測(cè)量植物DNA含量的主流方法。樣品制備的簡(jiǎn)便性和高樣品通量使其通常比其他方法（如福爾根密度測(cè)定法）更適合估計(jì)基因組大小、生殖多倍體水平、核復(fù)制狀態(tài)和內(nèi)多倍性（多聚體）。在這里，我們提出了四種樣品制備方案（完整細(xì)胞核的懸浮液），并描述了使用FCM分析核DNA量。我們考慮了必要的化學(xué)品和設(shè)備、測(cè)量過(guò)程、數(shù)據(jù)分析，并描述了植物材料遇到的最常見(jiàn)問(wèn)題，如次生代謝物的干擾。討論了內(nèi)外部標(biāo)準(zhǔn)化的目的和要求。強(qiáng)調(diào)了使用正確的術(shù)語(yǔ)來(lái)描述DNA數(shù)量和基因組大小的重要性，并解釋了其基本原理。
關(guān)鍵詞：
Sysmex Partec 倍性分析儀，多倍體誘導(dǎo)儀，靜水壓儀，CyFlow Ploidy Analyser 流式細(xì)胞儀，DNA含量測(cè)定
