Help! I got contaminants after electroporation - (Jun/29/2008 )
Hi everyone,
I am working a transformation with a different plasmid (I did lots before, but using a different vector). I am trying to get in-frame deletion mutants using pJRS233 derivatives. I got a perfect transformation using the vector only (as a test and optimization of conditions), but when I tried to use the in-frame deletion vector I got contaminants in my negative control (twice!!) I am trying to reduce all risk of contamination but it seems that is not working. Anyone with any ideas or similar experience?
Thanks a lot for your help.
-mgaleas-
What is the concentration of antibiotic you use for the plate? Sometimes it can be just the plate.
-timjim-
QUOTE (timjim @ Jun 30 2008, 03:16 AM)
What is the concentration of antibiotic you use for the plate? Sometimes it can be just the plate.
Today my boss told me that my technique was fine but we got contaminated soft agar so that was the cause (the soft agar didn't look contaminated when I used it, but I guess it was just a matter of some incubation). Thanks anyway.
-mgaleas-
QUOTE (mgaleas @ Jun 30 2008, 09:45 AM)
QUOTE (timjim @ Jun 30 2008, 03:16 AM)
What is the concentration of antibiotic you use for the plate? Sometimes it can be just the plate.
Today my boss told me that my technique was fine but we got contaminated soft agar so that was the cause (the soft agar didn't look contaminated when I used it, but I guess it was just a matter of some incubation). Thanks anyway.
Where do you live mgaleas? Sounds like someone I know
-cellcounter-
QUOTE (cellcounter @ Jun 30 2008, 01:12 PM)
QUOTE (mgaleas @ Jun 30 2008, 09:45 AM)
QUOTE (timjim @ Jun 30 2008, 03:16 AM)
What is the concentration of antibiotic you use for the plate? Sometimes it can be just the plate.
Today my boss told me that my technique was fine but we got contaminated soft agar so that was the cause (the soft agar didn't look contaminated when I used it, but I guess it was just a matter of some incubation). Thanks anyway.
Where do you live mgaleas? Sounds like someone I know
If that someone you know is a girl living in illinois and got contaminated electroporation plates, then it's me!
-mgaleas-
QUOTE (mgaleas @ Jun 30 2008, 04:15 PM)
If that someone you know is a girl living in illinois and got contaminated electroporation plates, then it's me!
That does narrow it down a bit, but a number would go a long way..
Seriously, I was thinking of someone down-under. Same initials.
-cellcounter-
QUOTE (cellcounter @ Jun 30 2008, 07:57 PM)
QUOTE (mgaleas @ Jun 30 2008, 04:15 PM)
If that someone you know is a girl living in illinois and got contaminated electroporation plates, then it's me!
That does narrow it down a bit, but a number would go a long way..
Seriously, I was thinking of someone down-under. Same initials.
I don't know many people (besides my family, of course) with Galeas as last name! If you happen to know me... well, don't tell people about my secret super powers. They're secret. 
-mgaleas-
QUOTE (mgaleas @ Jun 30 2008, 05:28 PM)
QUOTE (cellcounter @ Jun 30 2008, 07:57 PM)
QUOTE (mgaleas @ Jun 30 2008, 04:15 PM)
If that someone you know is a girl living in illinois and got contaminated electroporation plates, then it's me!
That does narrow it down a bit, but a number would go a long way..
Seriously, I was thinking of someone down-under. Same initials.
I don't know many people (besides my family, of course) with Galeas as last name! If you happen to know me... well, don't tell people about my secret super powers. They're secret. I can tell, telling sign was the cat,
shed the lad, I cried and wept.
Your secret is safe,
so long as mine is kept!
-cellcounter-