Advice on finding sonication parameters

Hello,

Thanks for reaching out. The sonication conditions we used were extremely mild. We performed sonication using a bath-type sonicator (Bioruptor®) for 5 minutes at low power settings (pulse: 30 seconds, rest: 60 seconds). This means a total of 4 pulses of 30 seconds each. We did not optimize these conditions but chose this duration and power based on previous experience with this machine. The objective was to disrupt nuclear membranes without using high concentrations of detergents. Since we used only 0.1% Triton X-100 in LSB1, we subjected the nuclei to mild sonication to achieve membrane disruption.

If you’re concerned that this sonication step could disrupt weak protein interactions, you could consider using a Dounce homogenizer instead. For us, the Bioruptor® was much more convenient.

I hope this helps.

Best regards,

Afzal