Hi everybody!
Today I saw a strange thing I have never seen before:
I ran a NuPage Novex 4-12% Bis-Tris Gel (Invitrogen) in 1x NuPage MOPS Buffer (diluted from 20x stock - 40 ml stock + 760 ml H2O) at 200V with constant voltage. When I returned to the gel after 30 minutes, the marker bands had migrated twice as far as usual, the buffer was boiling, the gel was cracking and the bottom of the gel had burnt!! Sample treatment and all conditions where the same as usual! Anaybody an idea what could be the reason for this overheating???
Hope you can help me!!!
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6 replies to this topic
#2
Adrian K
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Posted 31 January 2011 - 01:05 PM
Expired buffer? I don't know... my MES buffer works fine...
Did you check the amp?
Did you check the amp?
Expecting the world to treat you fairly because you are a good person is like expecting the lion not to attack you because you are a vegetarian.
..."best of our knowledge, as far as we know this had never been reported before, though I can't possible read all the published journals on earth, but by perform thorough search in google, the keywords did not match any documents"...
"what doesn't kill you, makes you stronger"---Goddess Casandra reminds me to be strong
"It's all just DNA. Do it."---phage434
..."best of our knowledge, as far as we know this had never been reported before, though I can't possible read all the published journals on earth, but by perform thorough search in google, the keywords did not match any documents"...
"what doesn't kill you, makes you stronger"---Goddess Casandra reminds me to be strong
"It's all just DNA. Do it."---phage434
#3
bob1
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Posted 31 January 2011 - 02:28 PM
It really sounds like your buffer is too concentrated, is it a new stock solution?
#4
HomeBrew
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Posted 31 January 2011 - 07:00 PM
Are you sure the power supply was delivering constant voltage? If it was set for constant current, the voltage would go high as the resistance increased (or you set it for 200 mA). In any event, if your apparatus was assembled correctly, and your buffer was correct, I'd try a different power supply...
#5
Hanna30
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Posted 01 February 2011 - 12:16 AM
Yes, I opened a new buffer stock.
Voltage was set constant to 200V.
Power supply worked normally the day before and the day afterwards - how's that possible
Voltage was set constant to 200V.
Power supply worked normally the day before and the day afterwards - how's that possible
#6
Adrian K
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Posted 01 February 2011 - 08:52 AM
Hanna30, on 01 February 2011 - 12:16 AM, said:
Yes, I opened a new buffer stock.
Voltage was set constant to 200V.
Power supply worked normally the day before and the day afterwards - how's that possible
Voltage was set constant to 200V.
Power supply worked normally the day before and the day afterwards - how's that possible
Hmn... the day before is ok, the day after is ok, but not the day itself....
Are you sure you are adding the correct stuff that particular day? Or maybe someone was sabotage your work.... is time for a hidden CCTV...
Expecting the world to treat you fairly because you are a good person is like expecting the lion not to attack you because you are a vegetarian.
..."best of our knowledge, as far as we know this had never been reported before, though I can't possible read all the published journals on earth, but by perform thorough search in google, the keywords did not match any documents"...
"what doesn't kill you, makes you stronger"---Goddess Casandra reminds me to be strong
"It's all just DNA. Do it."---phage434
..."best of our knowledge, as far as we know this had never been reported before, though I can't possible read all the published journals on earth, but by perform thorough search in google, the keywords did not match any documents"...
"what doesn't kill you, makes you stronger"---Goddess Casandra reminds me to be strong
"It's all just DNA. Do it."---phage434
#7
mdfenko
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Posted 02 February 2011 - 11:48 AM
is the hot gel from a different lot than the gels from the days before and after? it may have dried some, altered buffer, or be from a "bad" lot.
were the samples extra salty or in any way different from the samples you ran before?
(not to insult, but to troubleshoot) are you sure that you properly diluted the electrode buffer?
were the samples extra salty or in any way different from the samples you ran before?
(not to insult, but to troubleshoot) are you sure that you properly diluted the electrode buffer?
talent does what it can
genius does what it must
i do what i get paid to do
genius does what it must
i do what i get paid to do
