View source for Electroporation of C2C12 cells

== material ==
''Transfection''
* 1x Steinberger Medium
* 1x DMEM without serum

-	remove the old media
-	wash the cells with 5 ml PBS
-	remove the PBS
-	add 2 ml TE
-	put the cells nearly 4 min in the 37 ºC incubator
→ for 6 transfections

-	6 * 6 cm dishes
-	label those with the Plasmid name

-	cells from the incubator:
-	stop the TE with HS – media
-	resuspence the cells in the old media
-	remove the cells in the FALCON flask
-	spin the cells: 1000 rpm; 3 min

-	label the electroporation cuvetts: 1 – 6
-	after spinning: remove the old media in the waste
-	clean the pellet carefully with PBS ⇒ don’t  resuspend the cells
-	remove the PBS
-	resuspend the cells in 1,8 ml media (for each cuvette: 300 µl) (very well resuspending)
-	pipette in each cuvette 300 µl from the cellsuspendion
-	add in the cuvette the right plasmid
-	DNA – c.: 0,05 µg/ ml
-	closed he cuvette with the blue round top
→ Electroporation

-	room 434: the electroporator for the C2C12 cells
→ 130 V
→ pulse length: 30 ms
→ number of pulse: 5

1) empty cuvette with PBS → start (for cleaning)
2) pulse the cells

-	put the cells on ice
-	add in each cuvette 1 ml HS  (HS for the C2C12 cells) 

-	in every 6 cm dish: 3 ml HS