View source for Transfecting Cells

==Summary==
Cos 1 (African green monkey, from kidney cells). Cells are heterozygous, they all look different. We are transfecting Cos 1, 6 cm plates
* Make sure everything is clean and sterile 

===Materials===
*DNA, transfection lipofectamine 2000
* Antibiotic free DMEM (no serum), kept in the fridge 
* Sterile tips, tubes, pipettes (sprayed) 
* Make sure cells are 50-70% confluent 
* Work deep inside the hood


==Transfection mix (for this example)==
* Take sterile tubes for as many transfections you have, label
* In this example we needed six tubes
* Put 150 ul of media into tubes 
* You can reuse the tip if you don’t touch anything
* Make sure you don’t go over any open tubes or bottles, close tubes and bottles right away
* Into the 150 ul of media add about 1-2 ug of DNA (optimize DNA concentration first)
* Tap gently to mix
* Add 6.5 ul of lipofectamine (transfection media) 
* Pipette directly in the tube, flush, and tap
* re-cap / close
* Let it sit for 5-10 minutes (helps enhance transfection efficiency) 
* After optional 5-10 minutes, grab cell plates from incubator (careful not to open lid yet)
* Make sure tubes and plates are in order (1-6), set pipette to 170 (ish) ul, take up media from tube and add to plate (pipette at an angle so nothing falls in) spread media in drops in a circle 
* Give the dishes a gentle swirl, and place in incubator for a minimum of 24 hours and a maximum of three days. After 24 hours the DNa will be taken up, but after three days they will over grow and die