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293Flp cell transfection


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3 replies to this topic

#1 JdC09

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Posted 04 February 2009 - 08:32 AM

Hi,
Has anyone transfected cells with lipofectamine 2000, waited the required 4-6 hours and split the cells into a larger dish.
Would this affect transfection efficiency? The protocol says you can change the media after 4-6 hours which means you are effectively
taking off the liposomal reagents. So I was wondering if the cells can just be split at that time too...
Any advice would be very helpful.

Thanks

#2 scolix

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Posted 05 February 2009 - 07:48 AM

Transfection is stressful for cells and if they are split soon, many cell lines might not tolerate it well and you could have lots of cell death.

#3 cotchy

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Posted 06 February 2009 - 08:20 AM

it is recommended to wait up to 48 to 72 hours after removal of the lipofectamine to split the cells.

As scolix said they can be stressed and fragile after exposure to lipofectamine and so should be given 2-3 days to recover.

it does also depend on the cell type as some hardier cells such as hepG2 or similar can probably be trypsinised sooner, but if the transfection was to say immortalise primary cells or for fragile cells (where viabilty when passaging isnt 90 - 100 %) its best to give them time to recover

#4 cellcounter

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Posted 06 February 2009 - 08:32 AM

I agree with above responses. I am wondering why you want to do it?




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