Nucleotide analysis

Simon Simon
Fri Jan 21 14:42:42 EST 1994


>Does anyone know if perchloric acid lysis will work on yeast cells?  Will
>nucleotides be released from the cells or will the nucleotides be trapped
>by the cell wall?
>

Greetings buddies

Yeah, PCA extracts are great with yeast.  I use a lot more PCA than my 
mammalian cell colleagues, maybe to cope with the cell walls or something,
or maybe just historical accident. I use 5 ml of 3M PCA to lyse 4 g of cells.  
My cells are usually embedded in agarose threads for perifusion prior to
extraction, but it works fine on "loose" cells.  I usually snap-freeze the 
cells in liquid nitrogen and grind them up, allowing them to thaw in the 
presence of the PCA, and freeze-thaw them some more before proceeding with
the rest of the extraction prior to HPLC or NMR nucleotide quantitaion.

Best wishes

Simon Williams
Biochemistry
Cambridge, England



More information about the Yeast mailing list