Molecular biology sonicator Bioruptor® Plus

for DNA shearingfor RNA shearing

The Bioruptor® uses a unique system to uniformely process multiple samples in sealed tubes of 0.5 ml to 50 ml capacity. The built-in cooling system (water cooler and Single Cycle Valve) ensures high precision temperature control resulting in higher quality samples. An excellent device for shearing chromatin, cell and tissue disruption and many other applications (see below). To expedite the larger-scale phenotyping of pollen number difference within Arabidopsis family, we have modified an existing method more suitable for our purpose of pollen counting using a Bioruptor®. We grew four plants per genotype. Three flower buds per plant were harvested and dried at 65°C overnight. We collected flower buds with mature pollen, but before the anthers were opened (flower stage 13). We did not use the first and second flowers of the inflorescence, as they tend to show developmentally immature flower morphologies. 30 μl of 5% Tween-20 is added to the dried flower and samples are sonicated using the Bioruptor® Plus with high power mode for 10 cycles (sonication cycle: 30 sec ON, 30 sec OFF) to separate pollen grains from the anthers. Then, all pollen solutions are measured with a cell counter.