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DIAGENODE
BIORUPTOR
®
PLUS
USER MANUAL
Europe Diagenode sa
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diagenode.com
Efficient cell disruption with Bioruptor
®
Cell suspensions were sonicated for different periods of time ranging from 5 to 20 minutes. Two types of tubes were
tested: Diagenode’s 1.5 ml TPX tubes (M-50001) and Diagenode’s 10 ml tubes (AS-100). The efficiency of cell disruption
was initially determined by measuring optical density at 600 nm. The results indicated that the number of intact cells
decreases rapidly with increasing sonication time. After only 5 minutes of sonication, a significant number of cells
were disrupted (Fig.1). Similar results were observed using the Live/Dead BacLight kit (data not shown) which allows
the quantification of live cells with intact membranes and discrimination from cells with damaged membranes. Thus,
efficient cell disruption is observed after 5-10 minutes of sonication.
Figure 1: Effect of sonication on cell disruption
The number of intact cells after sonication was determined by measuring optical density at 600 nm. Optical density of
the cell culture before sonication (0 min) is arbitrarily set to 100%.
Sheared DNA is released during bacterial sonication
The disruption of bacterial cells by sonication releases DNA with maximum recovery after only 5 minutes of treatment
(Fig.2, A). The released DNA is fragmented with fragment size dependent on sonication time (Fig.2, B).
Figure 2 : Effect of sonication on DNA release
Figure A: The DNA concentration in each sample after sonication was quantified with the DNA BR assay kit (Invitrogen)
Figure B: An aliquot of each sample before (0 min) and after sonication was run in a 1.5% agarose gel stained with
SybrSafe and visualized in UV light. Lane M represents a 100 bp ladder.
B.
A.
M 0 min 5 min 10 min 15 min 20 min
