Haynes Lab:Notebook/Synthetic Biology and Bioinformatics for Predictable Control of Therapeutic Gene2/2012/10/29

From OpenWetWare

(Difference between revisions)
Jump to: navigation, search
(Autocreate 2012/10/29 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...
+
* Expand cell cultures: K-562, SK-N-SH
 +
 
 +
 
 +
----
 +
'''Expand cultures 4x'''<br>
 +
* Checked cells under scope. Good amount of cell division
 +
 
 +
* K-562 (suspension)
 +
** Split 10 mL culture into two (5 mL in 2 15 mL tubes)
 +
** Spun for 3 min, 1000 rpm, room temp
 +
** Resuspended each 5 mL pellet in 10 mL IMDM (10% FBS, 1% pen/strep)
 +
** Added 5 mL resuspension to 5mL medium in 75 cm flask (10 mL final volume)
 +
** Made 4 total 75 mL falsks
 +
 
 +
* SK-N-SH
 +
** Aspirate old medium, wash with 2 mL 1xPBS; detach with 0.5 mL Trypsin media (3 min.); collect in 3 mL EMEM (10% FBS, 1% pen/strep), add to 7 mL medium in 75 cm flask

Revision as of 15:48, 31 October 2012

Project name Main project page
Previous entry      Next entry

Entry title

  • Expand cell cultures: K-562, SK-N-SH



Expand cultures 4x

  • Checked cells under scope. Good amount of cell division
  • K-562 (suspension)
    • Split 10 mL culture into two (5 mL in 2 15 mL tubes)
    • Spun for 3 min, 1000 rpm, room temp
    • Resuspended each 5 mL pellet in 10 mL IMDM (10% FBS, 1% pen/strep)
    • Added 5 mL resuspension to 5mL medium in 75 cm flask (10 mL final volume)
    • Made 4 total 75 mL falsks
  • SK-N-SH
    • Aspirate old medium, wash with 2 mL 1xPBS; detach with 0.5 mL Trypsin media (3 min.); collect in 3 mL EMEM (10% FBS, 1% pen/strep), add to 7 mL medium in 75 cm flask


Personal tools