during resistance plasmid growth in transformation step, protocols usually say shake growth for 1-2 hr at 37 deg C after heat shock. when i grow it without shaking, i get colonies, when i grow with shaking, very very few colonies. although neither times i had any insert, but rest that for now. do you think i should grow stationary culture or shaking culture? and sometimes people say dh5alpha is very delicate and you need to keep on ice ass soon as its out of freezer and pipette gently to not kill the bacteria etc etc, which makes me think be ther reason why shake culture might be a problem but then they also say it is a very robust organism and so on. am confused! ! what is actually? and most importantly shake cuture or stationary?