96 filterplates to clarify E.coli lysates

Dear all,

I have used Nickel 96 expression screening protocols for years, our protocols are very robust and automated on a Tecan workstation. For the Ni96 protocols we are using crude lysates (from 1-4 ml cultures) on a 20 uM filter plate with 50 ul of beads because this works fine and the clarification in DW 24/DW96 is not trivial on a robotic plateform (if you are interrested by the details of our expression screening they are public). In theory I could scale this up a little but this might be the time to try something new, I might try more "expensive" columns like the Atoll from which I hear that if the lysate is not clarified I will quickly have difficulties.

To clarify by 96, I have used millipore filterplates often in the past but the one I used saturate around OD of 2, that's far too little. Some companies sell 2 ml filter plates that could do the job (my lysate volume is around 1 ml on average), does anyone have experience with clarification of lysate by filtration on a robotic platform at a scale of up to 4 ml with OD =20? I could of course tranfer the lysates to a centrifuge but that's not an option with the throughput that I want to reach.

Thanks for your help.



Posted on 19-Jun-2012 11:51 CEST
Renaud Vincentelli