Time course sonication experiment with the Bioruptor® PLUS using the buffers and protocol from Diagenode's Chromatin Shearing Optimization kit - Medium SDS
HeLa cells are fixed with 1% formaldehyde (for 8 min at RT). Nuclei isolation of 5 million cells are performed using buffers from Diagenode's chromatin Shearing Optimization kit - Medium SDS (Cat. No. AA-002-0100) and are then resuspended in 200 μl of Shearing Buffer S1 prior to chromatin shearing.
Samples are sheared during 1, 2, 3, 4, 5 or 6 rounds of 10 cycles of 30 sec ON/30 sec OFF with the Bioruptor® PLUS combined with the Bioruptor® Water cooler (Cat. No. BioAcc-cool) & Single Cycle Valve (Cat. No. VB-100-0001) at HIGH power setting (position H). All samples were treated with RNase prior to agarose gel analysis. 300 ng of each sample was analyzed on a 1.5% agarose gel (except for lanes 2 and 3).
In this example, the least time course condition corresponds to 3 rounds of 10 cycles (30 sec ON/30 sec OFF).
