1. Transform the cells with the recombinant plasmid and spread on LB agar plate
2. Pick up a colony in eg. 5 ml LB. Grow for few hrs
3. Use part of this culture eg. 0.5 ml for culture initiation in 100 ml LB.
4. And then go ahead till the desired OD achieved.
My doubt is for the step 2 and 3. Why are the cells initially grown in smaller amount of media? And then only part of the grown culture transferred to larger volume of media? Why not to grow the colony directly in 100 ml LB? Or why not to use whole culture obtained after step 2 for step 3?
Has anyone ever got different/negative results after doing so?
Edited by ram, 03 November 2009 - 06:42 AM.