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I too was waiting to hear replys to Peter Openshaw's request for info and
didn't see any so I will tell you some of our experiences. We also are using
a FACStar+ .Oddly enough we don't do anything different from other sorts
except to rinse the sheath tank out several times with PBS made with RNAase
free water (they call it DEPC whatever that is). We then fill the tank with
this stuff which the customer provides and run it through the fill and run
lines for an hour or so and that is it. I have a bottle of Ambion's RNAase
ZAP in the lab but don't even use it. The customer also brings over some
autoclaved ependorf tubes, sometime he coats them with calf serum or
doesn,t, and add a little of the PBS/DEPC to the bottom of them to sort into.
The operators used to be real paranoid and wear gloves when handling his
samples and when we saw he didn't, we stopped wearing them as well. We also
change the filter in the sheath tank during the rinsing step and keep the
sample tube and sorted cells cold (5-8 deg). We really don't talk much to
this fellow (don't like him) but he is able to isolate RNA from our sorts. If
you hear anything else that may be of interest please let me know. To me it
would be difficult to even estimate how much RNA it would be possible to
obtain so we have no quantitative data.
Sincerely, Marcia Woda
Manager, FACS lab UMMC
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