Help with DNA Quantification - Cell lysis for DNA quantification using Hoechst 33258 (Aug/20/2010 )
I need some help with how exactly to prepare my cells for quantification using Hoechst 33258 which I have with the Sigma DNA Quantitation Kit, Fluorescence Assay (#DNAQF).
I initially seed 50,000 cells in wells of a six well plate. I want to measure cell proliferation at different time points e.g. 72 hours, 7 days etc.
How exactly should I prepare the cells for use with this assay.... Theres so many protocols available for cell lysis but does anyone which one would suit best with this kit.
Any help would be very much appreciated
Lyse the cells in the plate - you could use freeze/thaw and then add the Hoescht 33258 in a detergent based buffer, leave for a time (1-2 hours should be heaps) and then read.
Great thanks for the help