Midiprep, vaccum to column

[biochembug]

Hi Folks

Has anybody applied vaccum to any commercially available columns at least for equilibration, binding and washing? Was wondering if we can cut short the time for plasmid prep!

Biochembug

[mdfenko]

are you asking about spin columns? or packed columns for lc?

you should be able to use a light vacuum with a spin column (too much and you may over compress the packing).

i wouldn't consider using vacuum with a packed column.

[biochembug]

Hey

My query is about the company supplied Midi or Maxiprep columns. We already have vaccum manifold for spin columns.

Biochembug

[mdfenko]

sorry, my mind blanked out the "midiprep" and just saw the "vacuum to column".

[relaxin]

Those midi and maxiprep columns are designed for gravity flow only. Any attempt to speed up the flow may result in poor recovery or quality of the plasmid. Just make sure the "supernatant" is free of debris before applying onto the column. Sometimes, debris or precipitate will pass through the syringe filter if you try hard to get the last drop of supernatant. When debris appeared in supernatant, you need to centrifuge them down.

[biochembug]

Hey Relaxin, your explain seems appropriate! I think I can give it a shot at lower psi.