Etchevers:Notebook/Genomics of hNCC/2009/03/09

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Freezing mistake

I left the cells at -20°C between Friday 5PM and Monday 12PM (67 hours). Thawed them out quickly and transferred into 50 mL warm DMEM + 12%FCS, spun down, resuspended each tube in 2 mL Rich medium, and replated 1 mL into 2 x 35 mm collagen dishes. Will see if there are any survivors.

Andrea Trentin has submitted a manuscript using 10 ng/mL EGF to get neurons rather than the 0.1 ng/mL I use currently. Made up the two B27 media to that concentration and replaced on the 4x flasks and on the coverslips. Refreshed the Rich media in both the R and S cultures; did not want to make up separate media. Will have to make up at least one more 100 mL aliquot though before freezing for good.

Received protocol from Alessandro for polysome preparation.