Immediately prior to use, the stock solutions were heated to 80°C for 10 minutes, followed by 10 minutes of sonication in a water bath sonicator (Branson model 2510, Fisher Scientific), and then mixed by high speed vortexing to ensure full dissolution of the glycolipids.
To maximize homogeneity, the particles were dispersed in the PDMS through 3 minutes of sonication with a probe tip and 10 minutes in a sonicator bath.
The first minutes of sonication increased the signal due to both increased DNA release and increased DNA fragmentation.
The optimal extraction conditions for tomato pastes (three minutes of sonication under 80 °C) were determined using the central composite design.
After 5-10 minutes of sonication, remove sample from bath and handshake again until sample is completely suspended (check visually).
During the formation process, a blue fresh product Cu(OH 2 is formed immediately after the addition of OH−, which turns into brown color of copper oxide after a few minutes of sonication.
Twenty minutes of sonication in dichloromethane was performed (Soniclean 160HT, 70 W, Soniclean Pty. Ltd., Thebarton, Australia) to transform any large sheets of pSi into smaller particles for recovery.
For in vitro assays, the 500 µM glycolipid stocks in DMSO were diluted directly into prewarmed (37°C) culture medium with vigorous mixing and 10 minutes of sonication.
Water and filters were extracted separately and filters containing planktonic material were extracted twice with 15 mL of methanol-acidified water (90∶10, v/v) by homogenizing for 1 2 minutes using a Polytron®, followed by 10 minutes of sonication in an ultrasonic bath.
Thus, ten minutes of sonication are sufficient to break significantly the GG chain backbones (the initial molecular weight was reduced to one third, see Section 3.3).
