Problem with RNA Preps from ARIA Sorts
scott.tighe at uvm.edu
Wed Jun 27 10:37:48 EDT 2007
We sort cells for RNA routinely with good success. I have attached a
flow sort for RNA protocol as well. Hope this document helps
Scott W. Tighe
Sr. Research Tech III
Vermont Cancer Center
University of Vermont
Microarray and Flow Cytometry Core Facilities
Health Science Research Facility 305
Burlington Vermont 05405
802-656-2557 -DNA Lab
Ck Poon wrote:
>I have a user who is sorting fresh PBMCs (from whole blood after ficoll) and
>fresh mouse splenocytes with at 70um nozzle at 70psi on an ARIA. He usually
>runs at a speed between 6k to 10k threshold events/sec.
>He is seeing a degradation of RNA preps from cells sorted on the ARIA
>compared to preps from cells sorted on a DiVantage.
>Here are some other pieces of information:
>- There are no UV lasers on both instruments.
>- Sorting at 25psi with a 70um nozzle on the DiVantage.
>- He is sorting into the same medium on both instruments.
>- Viability of the sorted cells with Trypan blue gave very good viability.
>Please ask if you need more specific answers.
>Thank you for any input anyone may have.
>Division of Experimental Medicine
>University of California, San Francisco
>UCSF Box 1234
>San Francisco, CA 94143-1234
>E-mail: cpoon at medsfgh.ucsf.edu
>Update Log: http://www.xanga.com/DEMFlowCore
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