Difference between revisions of "Preparation of High Serum for A1 cells"
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| Line 1: | Line 1: | ||
==== For 400 ml ==== | ==== For 400 ml ==== | ||
| − | FCS 40 ml | + | * FCS 40 ml |
| − | + | * Eagle’s MEM 252 ml | |
| − | Eagle’s MEM 252 ml | + | * DdH2O 100 ml |
| − | + | * Pen/ Strep 4 ml | |
| − | DdH2O 100 ml | + | * Glutamin 4 ml |
| − | + | * Insulin (stock: 1mg/ ml) 4 ml | |
| − | Pen/ Strep 4 ml | ||
| − | |||
| − | Glutamin 4 ml | ||
| − | |||
| − | Insulin (stock: 1mg/ ml) 4 ml | ||
* grow at 25ºC in 2% CO2 | * grow at 25ºC in 2% CO2 | ||
Revision as of 10:19, 13 February 2015
For 400 ml
- FCS 40 ml
- Eagle’s MEM 252 ml
- DdH2O 100 ml
- Pen/ Strep 4 ml
- Glutamin 4 ml
- Insulin (stock: 1mg/ ml) 4 ml
- grow at 25ºC in 2% CO2
- if you want to grow without CO2, then substitute L-15 for the MEM
- passage once per week 1:3
- wash with APBS (100 ml PBS + 25 ml ddH2O)
- trypsinize with TE
- stop with media
- centrifuge cells at 100 rpm 3 minutes
to make myotubes:
- wash cells 1x with APBS
- put cells in 0.5% FCSmedia
- wait 3 days