T7 Phage DNA isolation protocol

Dr. Duncan Clark news at hgmp.mrc.ac.uk
Fri May 4 02:55:51 EST 2001


In article <l03130300b717b67241e7@[130.95.224.182]>, the eminent
Stephanie Gunn at BIOSCI/MRC Human Genome Mapping Project Resource
Centre wrote
>Does anyone know of a relatively quick and simple
>method to isolate DNA from T7 bacteriophage?

Just like any other lytic phage. Check protocols for miniprepping
lambda. Single sheets formerly at the back of NAR used to be good for
quick protocols a few years ago.

The only real problem with T7 is that it will lyse your E.coli within 20
mins of infection so don't infect with too many phage at too low an OD.
If you can get the moi right (lot of luck/practice!) you can get in
excess of 30mg wt T7 DNA per litre.

Duncan 
-- 
Homogeneous Fluorescent Reporting Systems for Real-Time Quantitative PCR: 

Optimisation, Probe Technology & Future Systems

4-5 September 2001
King Alfred's College, Winchester, UK 

http://www.dera.gov.uk/html/news/events/real_time_quantitative_pcr.htm




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