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?
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.
More information about the Yeast