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!
0
1 votes
Changing the media after transfection of 293T cells
Started by distalless, Aug 16 2010 09:02 AM
7 replies to this topic
#2
laurequillo
-
- Active Members
-
- 265 posts
The Goddamn Batman!
1
Neutral
Neutral
Posted 16 August 2010 - 09:07 AM
distalless, on 16 August 2010 - 09:02 AM, said:
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!
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
"He must be very ignorant for he answers every question he is asked" Voltaire
"This is SPARTA!"
"I´m the goddamn batman"
"This is SPARTA!"
"I´m the goddamn batman"
#3
distalless
-
- Active Members
-
- 12 posts
member
0
Neutral
Neutral
Posted 16 August 2010 - 09:29 AM
laurequillo, on 16 August 2010 - 09:07 AM, said:
distalless, on 16 August 2010 - 09:02 AM, said:
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!
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
-
- Global Moderators
-
- 4,347 posts
Thelymitra pulchella
224
Excellent
Excellent
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
-
- Active Members
-
- 12 posts
member
0
Neutral
Neutral
Posted 16 August 2010 - 04:03 PM
bob1, on 16 August 2010 - 03:17 PM, said:
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
-
- Active Members
-
- 40 posts
Enthusiast
0
Neutral
Neutral
Posted 17 August 2010 - 12:11 AM
that's probably because your 293T's are over-confluent...not because of the transfectionmix
#7
distalless
-
- Active Members
-
- 12 posts
member
0
Neutral
Neutral
Posted 17 August 2010 - 10:38 AM
fysio lab, on 17 August 2010 - 12:11 AM, said:
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
-
- Active Members
-
- 40 posts
Enthusiast
0
Neutral
Neutral
Posted 26 August 2010 - 02:05 AM
distalless, on 17 August 2010 - 10:38 AM, said:
fysio lab, on 17 August 2010 - 12:11 AM, said:
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
