For plasmid DNA isolation, E.coli was incubated last night. But incubator doesn't work and I just knew it this morning. The incubator fixed now. Is it okay for my experiment?
Hi Timothy,
I think if your culture grew visually like normal or not so much less (you should not be able to look through the medium anymore), it should work. Also, plasmid preparation is not a big thing. If you don't get out much plasmid, you make a new overnight culture :)
Additional remark: If you're doing a real experiment (not just plasmid isolation) that should, for example, show how much plasmid you get out of your strain at different conditions, it is necessary to stick precisely to the conditions of your experiment!
Hi Timothy,
I think if your culture grew visually like normal or not so much less (you should not be able to look through the medium anymore), it should work. Also, plasmid preparation is not a big thing. If you don't get out much plasmid, you make a new overnight culture :)
Additional remark: If you're doing a real experiment (not just plasmid isolation) that should, for example, show how much plasmid you get out of your strain at different conditions, it is necessary to stick precisely to the conditions of your experiment!
Timothy An
As Sandra Wullich mentioned, innoculate new media with the previous culture and make new overnight culture. It would work just fine.
we found lower plasmid yields at 30ºC than at 37ºC, i would repeat the culture instead of wasting the miniprep kit, it's not a big deal.
Dear Timoty
As Sandra told you, if the culture growth well, it is ok.
The E.coli could be growth also at temperature lower than 37°C. The only differences is generally the growth rate slow down and you need longer time to acheive similar OD.
The only problem that you may have in your case, is that the cells grow less and you recover less DNA.
Did you measure the OD(600nm) of the culture?
Although plasmid replication is independent process rather than chromosom one, it seems lower temperature would impact on plasimd DNA concentration at RT as E.coli turbidity is low. You can easily re-do your experiment from scarch if you failed what you desired-not a big deal.
Just share your result with us what you have got from your plasmid extraction, which E.coli was grown at RT. Good luck
the cells should still grow at room temperature, only slower than at 37C. I would just allow them to grow for longer time.
Its okay........ You dont need to worry about it....
The E.coli could be growth also at temperature lower than 37°C. The only differences is generally the growth rate slow down and you need longer time to acheive similar OD. The only problem that you may have in your case, is that the cells grow less and you recover less DNA.
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