核DNA
福尔根染色
DNA
基因组大小
基因组
生物
样品(材料)
计算生物学
流式细胞术
计算机科学
生物系统
化学
色谱法
分子生物学
遗传学
基因
线粒体DNA
作者
Jaroslav Doležel,Johann Greilhuber,Jan Suda
出处
期刊:Nature Protocols
[Springer Nature]
日期:2007-09-01
卷期号:2 (9): 2233-2244
被引量:1198
标识
DOI:10.1038/nprot.2007.310
摘要
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.
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