SDS and acetate in DNA isolation

Austin P. So (Hae Jin) nobody at nowhere.com
Sat Oct 15 10:53:46 EST 2005


DK wrote:
> 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. 
> 
> DK

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 
the time).

http://biomed.humanapress.com/ChapterDetail.pasp?isbn=1-59259-409-3&ccode=1-59259-409-3:75

Austin







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