[RASMB] Procedures for Cleaning Ultracentrifuge Cells

[rkroe at rdg.boehringer-ingelheim.com]

Here is the general method that we use in Tom's lab:

 

To Clean AUC Cells:

1.      Remove housing plugs and red plug gaskets.

2.      Remove the solution from each side of the centerpiece using a
pipette and gel tips.

3.      Loosen the screw ring from the cell housing using the torque stand.

4.      Manually finish unscrewing the screw ring and remove the underlying
screw ring gasket.

5.      Turn cell housing upside down and push out the window holders and
centerpiece gently using a kim-wipe.

6.      Remove the windows, window liners, and window gaskets from the
window holders.

7.      Wipe the window liners, window gaskets, and window holders with
kim-wipes.  If any are excessively wet, rinse with water/5%
contrad/water/ethanol and dry.

8.      Wash the centerpieces and windows:

a.       Rinse with water.

b.      Rinse with 5-10% Contrad70 or RBS and either rub manually or use a
Q-tip to remove all protein residues from the windows and centerpieces.

c.       Rinse again with dionized water to remove all Contrad.

d.      Rinse with ethanol.

e.       Dry with Nitrogen.

 

When working with RNA do all of the above followed by (Carrie May's Method):

f.        Soak windows and centerpieces in RNAseZAP (Ambion cat# 9780.9782)
for 2 mins

g.       Rinse with DEPC treated water for 2 mins

h.       Rinse with ethanol for 2 mins

i.         Dry of nitrogen or let air dry on bench

 

 

-----Original Message-----
From: rasmb-admin at server1.bbri.org [mailto:rasmb-admin at server1.bbri.org] On
Behalf Of Allen H Pekar
Sent: Monday, February 21, 2005 10:25 AM
To: rasmb at server1.bbri.org
Subject: [RASMB] Procedures for Cleaning Ultracentrifuge Cells

 


RASMB Members: Tom Laue's e-mail of Feb. 14, 2005 states that he has seen
"surface effects (most frequently surface binding and aggregation)" .  He
also has seen "differences from window to window and centerpiece to
centerpiece (of the same kind), suggesting that cleanliness may be a
problem".  I contacted Tom about recommended cleaning procedures for
centrifuge cells and he will be sending me the methods used in his lab.  Tom
also suggested that I ask other RASMB members for their input.  If anyone
has a cleaning procedure which they think helps minimize differences between
cells, I would be very interested.  Thanks,  Allen Pekar  (Eli Lilly and
Company,  Indianapolis, Indiana) 

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