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

From OpenWetWare
Jump to: navigation, search
(Autocreate 2012/09/17 Entry for Haynes_Lab:Notebook/Synthetic_Biology_and_Bioinformatics_for_Predictable_Control_of_Therapeutic_Gene2)
 
(Entry title)
Line 7: Line 7:
 
<!-- ##### DO NOT edit above this line unless you know what you are doing. ##### -->
 
<!-- ##### DO NOT edit above this line unless you know what you are doing. ##### -->
 
==Entry title==
 
==Entry title==
* Insert content here...
+
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:
 +
 
 +
#Added thawed cells to 5 mL pure FBS in 15 mL conical tubes
 +
#Spun at 1000 rpm for 4 minutes
 +
#Aspirated off the FBS (to get rid of any toxins from the freezing solution)
 +
#Resuspended the cell pellet in 1mL 20% FBS/ 1% pen-strep DMEM media
 +
#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
 +
#HepG2
 +
#Luc #4
 +
#23;4;9 Luc EED
  
  

Revision as of 13:43, 31 October 2012

Owwnotebook icon.png Project name <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page
<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>

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