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culture time for maxi prep - (Mar/25/2006 )

i usually perform large prep after 14-18 hours culture. wht happen if i perform LP after 30 hours culture????

-T. reesei-

Just last week I did a miniprep of a 48 hr culture "just to try" and i got same results as a 16 hr culture. But theoretically after that time your cells will die, DNA be degraded and have less yield.

QUOTE (T. reesei @ Mar 26 2006, 04:59 AM)
i usually perform large prep after 14-18 hours culture. wht happen if i perform LP after 30 hours culture????

-tertu-

[after 30 hours culture i prepare DNA, i got good yeild but the purity is 1.2 biggrin.gif

i donno whether its for my unskilled manipulation or overculture time sad.gif

-T. reesei-

QUOTE (T. reesei @ Mar 27 2006, 04:04 AM)
[after 30 hours culture i prepare DNA, i got good yeild but the purity is 1.2 biggrin.gif

i donno whether its for my unskilled manipulation or overculture time sad.gif


Possibly a little of both. For a 500ml Maxiprep (kit), I culture overnight (18 hours max) and get good yield and purity. If your 260/280 is 1.2, I'd try cutting back on the culture time. Are you using a high or low copy plasmid?


b3ka

-b3ka-