Etchevers:Notebook/Genomics of hNCC/2009/06/05

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Premier passages cellules GRD d'EMB090003 et EMB090005

Cellules très confluentes sur la boite 60 cm (EMB090003, 8-9 sd) et la T25 (EMB090005, C17).

EMB090003 : counted 74 cells in one row, where 4 rows = x 104 cells/mL and had resuspended after trypsin - spindown into 0.5 mL Rich medium, made up last Friday or Monday. This is = 1.48 million cells.

  • Seeded 40% of these into a new collagen-coated T25 (= 592,000 cells or approximately 24K cells per cm2.
  • Froze remainder (888K cells) in 10% total DMSO in 0.6mL total Rich medium, currently in -80°C freezer.

EMB090005: counted 64 cells in one row, where 5 rows = x 104 cells/mL and had resuspended after trypsin - spindown into 0.5 mL Rich medium, made up last Friday or Monday. This is = 1.6 million cells.

  • Seeded 40% of these into a new collagen-coated T25 (= 640,000 cells or approximately 25.6K cells per cm2.
  • Froze remainder (960K cells) in 10% total DMSO in 0.6mL total Rich medium, currently in -80°C freezer.

How will you transfer these to liquid nitrogen? (Not a good idea to do tomorrow morning?!).

Spoke with M. Aubry from Leica Microsystems about service to our microscope and two dissecting scopes at Necker. 06 15 47 48 38.




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