Low copy protocol

Lynn Hughes lynn.hughes at jcu.edu.au
Mon Oct 7 18:07:12 EST 1996


Lou Cantolupo wrote:
> 
> Does anyone have a good protocol for isolating low copy # pDNA from
> bacteria?I have a method out of Maniatis - 
- grow 30ml of culture in 2YT with antibiotic, until late log phase 
(OD600 = 0.6)
- innoculuate 500ml 2YT broth with 25ml of above, use a 2 litre flask.  
Incubate for 2.5 hours at 37oC, shaking 300 cycles per min (OD 600 should 
be about 0.4)
- add 2.5ml of chloramphenicol (34mg/ml in EtOH), incubate for further 
12-16 hours, 37oC, shaking
- treat as normal maxiprep

Hope this is of some help

Lynn



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