Etchevers:Notebook/Genomics of hNCC/2009/03/06
|Genomics of human neural crest cells|| Main project page|
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Trypsinized with 10 mL freshly thawed trypsin the two 150 cm2 flasks - one in regular Rich medium (R), one in Rich medium prepared with Stemline Neural instead of DMEM/F12. The cells look the same overall.
Counts: S - 202 in 3 of 5 rows = 202 x 5 / 3 / 2 x 104 cells = 1,683,333 cells in 0.5 mL.
R - 246 x 5 / 3 / 2 x 104 cells = 2,050,000 cells.
Diluted each freezing volume with one volume of 20% DMSO in Rich medium. Placed in isopropanol freezer helper pre-cooled to 4°C, then at -20°C at 5PM.
The rest of the cells were re-pelleted, and resuspended in RLT + β-ME lysis buffer for RNA extraction, and placed in the freezer as well. If you can expect about 10 μg per 106 cells, here I can hope for 8 μg from the Rich cells and for 4.2 μg from the Stemline-Rich cells. These are in the culture room freezer for now with the serum in the bottom of the drawer.
Trypsinized the two neural differentiation formulations, diluted by 1/2. The counts were: