Doug 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
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