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Transfection Forum Transfection Forum. Cell transfection discussion including stable cells transfection, transient transfection. Troubleshooting and posting of discussions on lipofection, and reagents: Fugene, lipofectamine, and others.


detached 293T cells after addition of transfection reagent

detached 293T cells after addition of transfection reagent - Transfection Forum

detached 293T cells after addition of transfection reagent - Transfection Forum. Cell transfection discussion including stable cells transfection, transient transfection. Troubleshooting and posting of discussions on lipofection, and reagents: Fugene, lipofectamine, and others.


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Old 08-28-2010, 07:36 PM
Pipette Filler
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Default detached 293T cells after addition of transfection reagent



Hi there,

I'm new to transfections and I am not sure if this is suppose to happen in transfections. I am using HEK 293T cells (which are an adherent cell line) and when I seed them into 6-well plates 24 hours before I transfect and come in to start my transfection they are 50-60% confluent. I always carefully remove my media from the wells and when I add my transfection reagent-DNA mix I always add the reagent mix in a dropwise fashion-BUT after I do this I notice my cells clump up (ie round) and most of the cells are no longer adherent to the bottom of the wells. Is this going to affect my transfections? Will these cells adhere to the plate again within 24 hours?

I know 293 T cells are very easy to remove off plates from my experience...so I am wondering maybe I need to use fibronectin or something else so my cells don't come off after I add the transfection-DNA mix. Or does someone think there is something wrong with my transfection reagent? I am using TransIT-LT1.

Thanks for your help!
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