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maxi prep from mini DNA? - (Mar/22/2006 )

Hi all

When I want to prepare maxi prep transform mini prep DNA into bactria, pick a single colony to grow further. But colleque of my told me that I can directly grow the transformation mix on LB, without plating in between. What you think? Since I am not planning to sequence the maxi DNA it seems like a good idea. it will save a day for me.


Burcu smile.gif


works usually but:

1 you have much more change of contamination (all transformed cells must be correct so the slightest amount of 'wrong' DNA will be a problem)
2 unlike with the proper way (from one collony) the transformation mixture has many many non transformed bacteria. Amp does not kill bacteria and so when growing the big culture you may eventually get growth of the non transformed bacteria at some point and so get less yield.

if you are in a hurry and it is not for a major experiment you can try....



Amp is an antibiotic that will be destroyed by the resistant bacteria, so after a while, your amp concentration drops, which gives non-transformed bacteria a change to grow...