Abstrakt
In many fields of biodiversity research, nuclear DNA content is a crucial parameter of the study organism (individual, cellular type), allowing, for example, ploidy determination, cell-cycle analysis or selecting suitable organisms and optimal strategy for whole genome sequencing (WGS). Due to lower sequencing costs, small genome size represents a major advantage for WGS projects.
Not surprisingly, most DNA content estimates available for small genomes have been derived from WGS data. On the other hand, the routine use of WGS as a method for genome size estimation has been discouraged due to its poor quantification of genomic content of repetitive elements (e.g., present in centromeres or telomeres) that may significantly underestimate the true DNA amount.
Currently, the most suitable method for the task is flow cytometry (FCM), a rapid and easy to perform technique, using which the DNA content is estimated from the mean fluorescence intensity of nucleic acid binding dye (e.g., propidium iodide, ethidium bromide). The FCM is routinely used in immunology, cancer research or plant and animal studies, however, its application on organisms with small genomes can be highly challenging.