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Working with F plasmids


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#1 pDNA

pDNA

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Posted 28 May 2010 - 08:10 AM

Dear people,

i'm currently trying to work with a mini-F plasmid (never worked with F plasmids before).
Are there some things one has to consider when working with this single copy plasmids?
For example:
1) Are there special protocols available for the isolation of F plasmids? ...or is it sufficient to use spin columns and use the low copy number protocols?

2) Are there special E. coli strains needed for successful cloning with F plasmids? There are some strains that do already contain a F plasmid (like JM109 for example) ...is it possible that E. coli contains two F plasmids? ...what will happen? (we accidentially used JM109 for cloning of a fragment into this F-mini plasmid and got weired results ...therefore i'm interested if this will be a problem due to recombination or if the strange problems are due to some other issue).

Many thanks in advance!
Your suggestions are highly appreciated!

Regards,
p




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