# Disaggregate mechanically ## Vortex with glass beads

Protocol: 5 mL sample + 1 mL [3 mm glass beads](/encyclopedia/item-register/consumables/glass-beads-3-mm-50-g.md). 3 min at max speed. ## Bead beating

Protocol: 5 mL sample + 1 mL [3 mm glass beads](/encyclopedia/item-register/consumables/glass-beads-3-mm-50-g.md). 1 min at max speed. ## Ultrasound bath

* Use 37-80 KHz cavitation frequency as lower frequency ultrasound (typically 20khz) results in aggressive cavitation that may induce lysis. * Use glass vessels for optimal propagation of ultrasound as the conventional plastic test tubes dampens ultrasound. * Make sure ultrasound bath is clean and that vessel is placing at the recommended water level and position in the bath * Typical protocol: 5 mL sample, 5 min @40 KHz in a glass vessel. If the ultrasound bath is equipped with a sweep functionality, use this. ## Ultrasound probe

* Typically, the rod-type sonicator probes are more aggressive than the above bath types, but amplitude, wattage and frequency parameters can be tailored. * Use [probes for small volumes](https://bandelin.com/en/shop/sonopulse-ultrasonic-homogenisers/sonopuls-hd-5020-homogeniser/) and higher frequencies than 20khz. * Cool sample in ice bath when performing homogenization * Typical protocol: 1mL sample on ice. Example: 10 cycles of 30 sec pulses at 20 watts, with 30 sec idle time. ## Rotor/stator blender

* Handheld rotor/stator equipment, e.g. [tissueruptor](https://www.qiagen.com/at/spotlight-pages/ias/automated-qpcr-workflow/detection/tissueruptor-ii/) or ultra-turrax types. Preferentially choose disposable probes to minimize contamination * Walk-away platforms like the [tube drive](https://www.ika.com/en/Products-Lab-Eq/Dispersers-Homogenizer-csp-177/ULTRA-TURRAX-Tube-Drive-cpdt-3646000/) and the DT-20M tubes are sealed to minimize aerosol formation. * Add antifoam agents, e.g. 1 µL Sigma antifoam 204 to 5 mL of culture * Typical protocol: Add 0.005-0.01% antifoam. 5-10mL sample. Homogenize for e.g. 2-5 min at max speed. ## Syringe homogenization

Protocol: Passage 200 µL sample through 27G cannula 20 times. --- # Agent Instructions: Querying This Documentation If you need additional information that is not directly available in this page, you can query the documentation dynamically by asking a question. Perform an HTTP GET request on the current page URL with the `ask` query parameter: ``` GET https://help.sbtinstruments.com/protocols/prepare-your-sample/break-up-clumps-and-chains/disaggregate-mechanically.md?ask= ``` The question should be specific, self-contained, and written in natural language. The response will contain a direct answer to the question and relevant excerpts and sources from the documentation. Use this mechanism when the answer is not explicitly present in the current page, you need clarification or additional context, or you want to retrieve related documentation sections.