Plasmid Extraction from a big volume of pellet - Molecular biology (Mar/20/2006 )
I need a protocol for amplifying a library. I have a big quantity of cells (pellet of 45 gr), but i dont know if it works with QUIAGEN kits or with some maniatis protocols.
Thank you for your help.
best choice is to do an alkaline lysis.
Measure weight of a 50mlculture pellet.
You need 2.5ml of buffer 1, 2 and 3 for that weight.
Adjust protocol for your pellet.
kits will be saturated and you won't be able to recover your plasmid.
If you decide to use a kit that can handle a large prep (Maxiprep, etc.), you still need to be careful not to overload the columns. Split up your pellet when resuspended in Resuspension buffer. And make sure your sample is completely lysed and not thick and dark before loading onto the column.