Protocol describes the quantitation of DNA using Hoechst 33258, a fluorescent dye that binds to double-stranded DNA. Fluorometry is simple and more sensitive than spectrophotometry, and allows the detection of nanogram quantities of DNA. The assay can only be used to measure the concentration of DNAs whose sizes exceed ~1 kb, as Hoechst 33258 binds poorly to smaller DNA fragments. - [Read Fluorometric Quantitation of DNA Using Hoechst 33258 Protocol]
In this protocol, bacterial cells are lysed by being subjected to short, intense treatments with ultrasound, which breaks the cell walls and shears the DNA into sizes that will not affect the viscosity of the samples. Note that this method causes some denaturation of the samples. The resulting lysate is ready for preclearing. - [Read Immunoprecipitation: Lysing Bacteria by Sonication Protocol]
Feeding euplotids with algae can lead to asynchronous cell starvation and vastly different cell sizes within a culture. Asynchronous starvation also leads to different levels of mating competence. Furthermore, algal pigment remnants can interfere with many applications (e.g., fluorescence microscopy). - [Read Refeeding Marine Euplotids with Bacteria Protocol]