Haynes Lab:Notebook/Synthetic Biology and Bioinformatics for Predictable Control of Therapeutic Gene2/2012/09/17

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==Entry title==
==Entry title==
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The previous attempt to get the cells started officially did not work. The cells did not attach and were floating, even after three days of incubation at 37C. I did another thaw and this time I did the following:
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#Added thawed cells to 5 mL pure FBS in 15 mL conical tubes
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#Spun at 1000 rpm for 4 minutes
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#Aspirated off the FBS (to get rid of any toxins from the freezing solution)
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#Resuspended the cell pellet in 1mL 20% FBS/ 1% pen-strep DMEM media
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#Added the resuspended cells to 3 mL of 20% FBS/ 1% pen-strep DMEM media in a 6-well culture dish (one cell line per well). This gives the cells less surface area to stick to and crowds them together, which sometimes promotes cell growth.
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The cell lines I thawed are
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#HepG2
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#Luc #4
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#23;4;9 Luc EED

Revision as of 16:43, 31 October 2012

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Entry title

The previous attempt to get the cells started officially did not work. The cells did not attach and were floating, even after three days of incubation at 37C. I did another thaw and this time I did the following:

  1. Added thawed cells to 5 mL pure FBS in 15 mL conical tubes
  2. Spun at 1000 rpm for 4 minutes
  3. Aspirated off the FBS (to get rid of any toxins from the freezing solution)
  4. Resuspended the cell pellet in 1mL 20% FBS/ 1% pen-strep DMEM media
  5. Added the resuspended cells to 3 mL of 20% FBS/ 1% pen-strep DMEM media in a 6-well culture dish (one cell line per well). This gives the cells less surface area to stick to and crowds them together, which sometimes promotes cell growth.

The cell lines I thawed are

  1. HepG2
  2. Luc #4
  3. 23;4;9 Luc EED


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