KT 01/05/06
Bringing up Frozen HT22 Cells
1. Make sure you have HT22 media ready and warm
a. 40mL FBS
b. 1mL P/S
c. 460mL DMEM w/GLUT
2. Look in liquid nitrogen folder in Levitt lab computer room to locate the rack
and box number of the cells you want to bring up
3. When opening liquid nitrogen tank locate the rack and box number
4. Remove the cells from appropriate rack and box
5. Go to liquid nitrogen folder and mark that those cells have been removed
from rack and box
6. Put the vials of cells into a bucket of dry ice and bring to waterbath
7. Hold tube with cells in water bath for approximately 1-2min, until cells
have thawed
8. Add 0.5-1mL of HT22 media to cell suspension
9. Mix gently
10. Add 10mL of HT22 media to a flask
11. Add cells and media mixture to flask
12. Let grow overnight and check growth the following morning to determine
when to split