transfection.. - help needed (Nov/29/2005 )
This is regarding HeLa cells. before the transfection, cells are in DMEM with high glucose+penicillin+streptamycin+10%serum (as per the protocol)
My question is ,
1. how does high/low concentration of glucose effect cell growth/transfection ?
2. Right after transfection, shall i add the same medium or just DMEM with high glucose ( i have already purchased DMEM with high glucose and so i dont have just DMEM) ?
3. how should the media be aliquoted ???? i was planning to add the required amount of strp/pen and 10% serum to the original 500 ml bottle and then aliquot in 50 ml falcons.
,,,,,,,,,,,,,,,is this okk or there is another way,,,please let me know..
As people grow hela's in high/low glucose, I don't think it matters too much. Maybe glucose makes them grow a little faster because of higher energy contents?
There's another topic that deals with Hela transfection http://www.protocol-online.org/forums/inde...showtopic=10960. I don't think that the glucose will have much effect, after transfection, wash your cells with pbs and add fresh complete medium (DMEM (high gluc or not) + 10%FCS + P/S).
As I'm using a lot of media, I just take a 500 ml bottle, add my FCS and P/S and store it at 4°C. But I never have the bottle longer than 2 weeks.
cells should be maintained in same gluc conc before and after transfection, to be sure of the effects of what you're transfecting. Beyond this point, high or low glucose in transfection... i don't know the effect.
For preservation of medium : i add FCS P/S and store the bottle at 4. Generally used before getting the "old" age of 2weeks