Lysing yeast spheroplasts

Dan Zabetakis dan at cubsps.bio.columbia.edu
Thu Jul 13 23:59:59 EST 1995


>In Article <3u1cjb$5et at seymour.sfu.ca>, wasylnka at sfu.ca (Julie Wasylnka) wrote:
>
>>I am trying to lyse yeast spheroplasts in order to make yeast DNA.  I 
>>have a protocol which used DTT. I was wondering if I could substitute 2-ME 
>>for DTT and what concentration I should use? (The protocol suggests 1.8 
>>mM final DTT).

  I think the DTT or 2-ME is for the lysing enzyme rather than the lysing
itself. Doesn't your procedure say to wash the spheroplasts prior to
lysing?  Get's rid of the stench. But you can substitute 2-ME for lyticase
at least, if that's what you are using. 1mM should be fine.

DanZ

  
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