New paper on Nature Protocols

The Central Europe plant flow cytometry triangle team, i.e., Jaroslav Dolezel (Olomouc), Johann Greilhuber (Vienna) and Jan Suda (Prague), has just published another very interesting manuscript on the estimation of nuclear DNA content in plants using cytometry. This review and step-by-step protocol has just been published in Nature Protocols the online resource for protocols of the Nature Publishing Group.

The article is very carefully written and presents step-by-step protocols for the analysis of nuclear DNA content in plant tissues, with the underlining of the procedure's critical steps. A very useful troubleshooting table is also provided, with the possible reasons and solutions for the most common problems that may appear upon analysis of plant tissues using flow cytometry. Therefore the article is utterly recommended not only for beginners but also for more experienced users of the area of plant FCM.

Below follows the abstract and the link to the online version of the manuscript.

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.

Author for correspondence: Jaroslav Dolezel; e-mail: dolezel@ueb.cas.cz

Online version