cell lysis after induction - (Sep/30/2009 )
I've been looking all over the web for information on this and I was wondering if someone could shed some light for me. In a system manual that came with my plasmid (pTriEX), it describes how to do a total cell lysis using Laemmli's buffer--but for 1mL of sample (100uL of Laemmli buffer per 1mL of culture). I was wondering if it was ok to scale up proportionally to 40mL? Is there a max amount of Laemmli buffer that one can use...?
The lysis will be based roughly on an area or cell confluency estimate for the protocol. 100 ul per well of a 6 well plate is probably about the volume you need. Using too much lysis buffer will mean that your samples will be too dilute to use for your end application (e.g. western blot).