painful gel loading???!!!!!!
David L. Haviland, Ph.D.
dhavilan at IMM2.IMM.UTH.TMC.EDU
Thu Jan 30 00:24:15 EST 1997
At 12:05 1/29/97 -0500, Darren Natale wrote:
>xudong huang wrote:
>> I have a painful problem about agarose gel loading. The DNA samples do
>> not stay at the bottom of wells, intead, they float out of wells and
>> diffuse into the buffer. The loading buffer I use is 6XBPB, the gel
>> running buffer is 1X TBE. It is not because of air bubbles or oil
>I had the same experience once. In my case, I suspected the problem was
>due to the running buffer (for whatever reason), since I re-use it
>several times. The problem disappeared when I re-loaded the same
>samples onto to a fresh buffered gel. Go figure.
Interesting! I've run a TAE buffer upwards of 5-7 times without noticing
an effect on the samples. What hasn't been discussed is the composition of
the loading buffer of the original post. A 6X BPB is 6X in what? I use
a TAE 1X that is 15% Ficoll, 10mM EDTA, and either BPB or XC are added
sparingly to color.
Where I have had sample leave the wells was due to either ethanol,
isopropanol, mineral oil and the best for "tornadically" removing one's
samples from wells was phenol. I would suspect one of these.
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