0.1mg/ml DNase type IIS from Sigma added to the pellet following
thawing should solve your problems. Make sure there are some
divalent cations in your buffer. The treatment is repeated during
processing if clumping reoccurs and we always add DNase to the sort
tube to prevent any problems during sorting - I would avoid
extensive vortexing as this may provoke more cell damage. Also
strain through 70uM mesh to remove non-DNA clumps.
Best of luck
################################################################
From: Dr Richard Darley, Ph.D.
LRF Differentiation Unit
Department of Haematology (A7)
University of Wales College of Medicine
Heath Park
Cardiff, CF4 4XN
U.K.
tel (+44) 12 22 74 34 85
or (+44) 12 22 74 45 24
Fax (+44) 12 22 74 45 23
e-mail: darley@cf.ac.uk