Our lab uses Bead Ruptor 12 ( Omni [Kennesaw, GA] ) to homogenize biofilms so we can measure CFU. For a while, we processed the samples at the high setting for 99 sec (the max settings). I recently tried optimizing this and found that beating them twice for 198 sec (beating twice in a row for 99 sec), gave slightly more CFU (not even a two-fold difference). However, now that I am using this processing time, I find myself getting more inconsistent results and slightly more leaking, which I had never dealt with before. When I optimized the protocol, I did not beat the samples consecutively like I am doing now. Does anybody know if beating samples consecutively (i.e. beating for 99 sec twice in a row) could do anything to the bead beater tubes that make them more prone to leakage?
Hi Sakila,
It is often recommended to have rests in between cycles of bead beating, to allow the sample and the machine to cool down.
While I have not previously used a Bead rupture, with FastPrep, Ribolyser and bedbug machines I have only ever used for 30-45 seconds before allowing the samples to cool on ice and also allowing the machine to cool for a short while.
I imagine this could also help reduce the stress on your tubes, preventing leakage.
It may also be the tubes themselves, do these tubes contain an o-ring to seal the lid very tightly?
If not, it may be worth trying to seal them with parafilm as an extra layer of protection.
Hope this is helpful,
Eleanor
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