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Changing the media after transfection of 293T cells


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#1 distalless

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Posted 16 August 2010 - 09:02 AM

So I transfected 293T cells with 5 different plasmids on Saturday. I was supposed to change the media on Sunday, but the door to the tissue culture room was locked and I could not get in.

Do you think the cells will still be OK and the transfection will work? Thank you!

#2 laurequillo

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Posted 16 August 2010 - 09:07 AM

So I transfected 293T cells with 5 different plasmids on Saturday. I was supposed to change the media on Sunday, but the door to the tissue culture room was locked and I could not get in.

Do you think the cells will still be OK and the transfection will work? Thank you!


293t are quite tough, so if they are still alive ( I dont know what kind of reagent did you use, and what is the toxicity) I would continue with the experiment
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#3 distalless

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Posted 16 August 2010 - 09:29 AM


So I transfected 293T cells with 5 different plasmids on Saturday. I was supposed to change the media on Sunday, but the door to the tissue culture room was locked and I could not get in.

Do you think the cells will still be OK and the transfection will work? Thank you!


293t are quite tough, so if they are still alive ( I dont know what kind of reagent did you use, and what is the toxicity) I would continue with the experiment


I used Lipofectamine 2000. How would I be able to tell if they are still alive? The plate seems very confluent.

#4 bob1

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Posted 16 August 2010 - 03:17 PM

If the plate is confluent, the cells are probably alive - dead cells are usually rounded and floating.

#5 distalless

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Posted 16 August 2010 - 04:03 PM

If the plate is confluent, the cells are probably alive - dead cells are usually rounded and floating.


Yes the plate is confluent, but the cells are coming off the plate. In fact more than half of the cells have peeled off the plate!

#6 fysio lab

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Posted 17 August 2010 - 12:11 AM

that's probably because your 293T's are over-confluent...not because of the transfectionmix

#7 distalless

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Posted 17 August 2010 - 10:38 AM

that's probably because your 293T's are over-confluent...not because of the transfectionmix


Do you think that I can still go ahead and use the supernatant for viral transduction? Thanks!

#8 fysio lab

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Posted 26 August 2010 - 02:05 AM


that's probably because your 293T's are over-confluent...not because of the transfectionmix


Do you think that I can still go ahead and use the supernatant for viral transduction? Thanks!


i think so, but pass the medium over a 0.45 filter to get rid of the dead cells




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