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
Cambridge, England

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