Resuspend the cells to be crushed in suitable buffer, try to keep the volume to less than 25 mls as this will then all go into the cell in one go.
Put a spot of glycerol onto the rubber seals then put the plunger and lid into the cell, also fit the outlet valve, with a new nylon ball in place, and both the inlet and outlet pipes with some tubing and leave them open.
Place the cell into the press, close the clamp across the cell to lock it in place and finger tighten the two screws to keep the cell from rising up during filling.
Turn back the pressure regulator and switch the selector on to High.
Turn on the press and wind up the pressure to around 1000 to 1100 on the dial. The press will need to rise up and engage with the top of the cell before you can set this. Then fit the refill clamp to the top of the press.(may need lowering a bit to get it on)
OPTIONAL Add 240µl M MgCl2, 24µl M MnCl2 and 10 - 20µl DNase to the sample.
Open the inlet valve gently and place into the sample. Close the outlet valve then set onto Down - the sample will be drawn up into the cell. When it is full close the inlet valve.
To crush the cells set to High and let the pressure build up, the gently open the outlet valve so that the crushed cells flow slowly enough that the pressure doesn't fall significantly - to ensure adequate lysis.
To refill close the outlet valve, open the inlet valve and follow from (7) above.
Clean up and dry the cell thoroughly before returning to G6.
Notes: Pressure setting on the gauge of 1000=3000psi in the cell on the medium setting, 18,000 psi on the high setting
I usually send my cells through twice to ensure good lysis.