Heriberto Cerutti, a post-doc in the Boynton-Gillham lab, tells me that
eliminating the final high-speed centrifugation prior to freezing the
autolysin prep seems to result in higher activity. He thinks perhaps the
enzyme is partly bound to cell debris. Since freezing at -80C would
presumably kill any surviving Chlamydomonas cells in any case (as everyone
who's tried to preserve Chlamy by freezing can attest!), this step would
seem to be non-essential. Has anyone else experimented with this?
chlamy at acpub.duke.edu