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Plasmid Miniprep

Plasmid Miniprep

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2 replies to this topic

#1 Epigeneticist

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Posted 19 November 2012 - 05:57 PM

Growing bacteria for plasmid minipreps usually requires a 12-16hr incubation. Longer incubation can cause cell lysis and loss of plasmid. Too short of incubation means less growth and less plasmid. Sometimes I don't have time to miniprep when the bacteria are ready, especially when I have 20 samples or so. Is there a good stopping point? I was wondering if I could remove my culture at 12-16 hours, pellet the cells, aspirate the media and refrigerate or freeze the pellet until I have time to do the miniprep? I thought this would be ok as long as the media is completely removed otherwise the cells would lyse during the freeze-thaw and release plasmid which would be lost during aspirating.

#2 leelee

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Posted 19 November 2012 - 06:32 PM

I've done what you just described (pellet, aspirate media, freeze), and the subsequent mini-prep worked just fine.

#3 Trof

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Posted 20 November 2012 - 03:08 PM

Yes, common method for keeping bacterial pelets before plasmid isolation. OK for (at least) weeks. Let the bacteria thaw directly by adding lysis solution and mixing after that.

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