SDS and acetate in DNA isolation
Austin P. So (Hae Jin)
nobody at nowhere.com
Sat Oct 15 10:53:46 EST 2005
> In article <dippks$a7a$1 at nntp.itservices.ubc.ca>, "Austin P. So (Hae Jin)" <nobody at nowhere.com> wrote:
>>1. punch holes in the cell wall. The idea is that timed degradation
>>(NaOH or lysozyme) will create a "cage" in the peptidoglycan wall.
>>Eventually, the holes are large enough to let out plasmid, but small
>>emough that genomic DNA is retained within the "cage". Too long of an
>>incubation leads to release of genomic DNA, too little leaves a lower
>>yield of plasmid.
> I don't think this is correct. Your description makes it sound as if cell
> wall remains but is full of holes. That is clearly not the case - the cells
> get completely lyzed, with no structure remaining. Which is quite
> obvious from the fact that the strongly light scattering suspension
> turns into almost clear solution.
Hmmm...obviously I have put my foot in my mouth and given credit where
it was not due! I suppose it is worse promulgating false information...
I was mistaken about the alkaline lysis prep (I use the boiling prep all
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