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Just to update. I switched to old school plasmid preps. Using isopropanol and 70% ethanol to clean my plasmids. Avg of ~1ug of DNA 260/280 ~1.7-1.9
I think our kit is probably off, pH or something. Glad I worked it out! Thanks for the suggestions.
D
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In Topic: Clean genomic DNA and plasmid DNA extraction from E.coli
10 January 2013 - 03:26 PM
Hi I just wanted to jump in and ask another question:
In general when doing a gDNA prep do plasmids get pulled down with it?
Specifically we have a low copy plasmid that we are trying to use for homologous recombination
and we are trying to screen for the first recombination event. I suppose if a cross over has occured
the plasmid backbone will be attached to gDNA. However if recom. has not occured we could still get
non-specific amp if plasmid gets pulled down in the gDNA prep.
Hope I haven't confused
D
In general when doing a gDNA prep do plasmids get pulled down with it?
Specifically we have a low copy plasmid that we are trying to use for homologous recombination
and we are trying to screen for the first recombination event. I suppose if a cross over has occured
the plasmid backbone will be attached to gDNA. However if recom. has not occured we could still get
non-specific amp if plasmid gets pulled down in the gDNA prep.
Hope I haven't confused
D
