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TY - JOUR AU - Doležel, Jaroslav AU - Greilhuber, Johann AU - Suda, Jan PY - 2007 DA - 2007/09/01 TI - Estimation of nuclear DNA content in plants using flow cytometry JO - Nature Protocols SP - 2233 EP - 2244 VL - 2 IS - 9 AB - 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. SN - 1750-2799 UR - https://doi.org/10.1038/nprot.2007.310 DO - 10.1038/nprot.2007.310 ID - Doležel2007 ER -