MR600D Standardized Cell Disruption Workflow

MR600D Standardized Cell Disruption Workflow

1. Purpose and Principle

Cell disruption is a core sample-preparation step for protein extraction, nucleic acid extraction, enzyme activity assays, cytokine analysis, and metabolite workflows. The MR600D non-contact ultrasonic disruptor uses liquid-phase cavitation to generate uniform microjets and shear force, disrupting cell membranes and cytoskeletal structures without direct probe contact.

2. Sample and Reagent Requirements

• Sample types: adherent cells, suspension cells, primary cells, stem cells, and other mammalian cells.
• Cell amount: typically 1 x 10^6 to 1 x 10^7 cells per sample, adjusted by cell type and downstream assay.
• Buffers: RIPA buffer for protein extraction, sterile lysis buffer for nucleic acid extraction, PBS, and fresh protease or phosphatase inhibitors when needed.
• Device condition: MR600D pre-cooled for about 20 min with stable 4 C water circulation.

3. Standard Workflow

1. Collect adherent cells by digestion and PBS washing, or collect suspension cells by centrifugation.
2. Resuspend the pellet in 200-500 ul suitable lysis buffer and keep samples on ice for 5-10 min.
3. Briefly centrifuge to collect liquid and remove bubbles before sonication.
4. Place tubes symmetrically in the MR600D 1.5 ml rack.
5. Select the standard or gentle disruption condition and process under low temperature.
6. Centrifuge at 12000 rpm, 4 C for 10 min and collect the supernatant for downstream assays.

4. Reference Conditions

Sample type	Amplitude	Pulse mode	Total time	Target
Common cell lines such as HEK293, Hela, and HepG2	30%	5 s on / 10 s off	120-180 s	Complete lysis with uniform turbidity
Sensitive cells such as stem cells, primary cells, and immune cells	30%	3 s on / 7 s off	90-120 s	Gentle lysis with activity retention

5. QC and Troubleshooting

Qualified lysate should be uniform, without visible cell clumps or heavy precipitation. Microscopy can confirm the absence of intact cells. For incomplete disruption, extend total time or improve resuspension. For over-disruption, reduce total time and keep samples colder. For precipitation, verify buffer compatibility and centrifuge before downstream assays.

6. Summary

MR600D provides a low-temperature, non-contact, repeatable route for cell disruption. It reduces probe contamination and local overheating while supporting protein, nucleic acid, and activity-preserving workflows.