Inoue help - again...
s535290 at aix1.uottawa.ca
Tue Feb 3 09:29:32 EST 1998
sorry to post again...had forgotten something else...
just some quick help. I'm going to try making competent cells the inoue
way, and only had the recipe for making the cells, but not for the actual
transformation. Do I just do the transformation the "regular" way ?
ie.thaw on ice, add DNA, wait 20-30 minutes, heatshock, chill, add SOC
and grow at 37 deg for 60 minutes, plate on selective media.
I also wanted to ask what to use to bring the pH of the transformation
buffer up to 6.7. I tried KOH and that didn't go well at all...I got some
sort of precipitate (I guess some Manganese salt, since it has a purplish
color). Should I use diluted KOH as opposed to 1 N KOH ?
thanks in advance,
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