Subject: PFGE trouble

From: [email-masked] (Dunkles group)
Date: 26 Jan 1996 13:19:45 GMT

My problem is that I am getting streaks instead of bands.  I believe that
this is related to the fact that my buffer is giving slightly lower mA
readings (18 instead of 21; 15 instead of 18) than usual.  

I have re-made the buffer exactly the same way that I always do when I get
high quality results, and it gives the same reading.  I sent the power
source in for repair, as I did 6 months ago when the same thing happened,
and they found the the readings were slightly HIGH and re-calibrated the
power supply (but THAT, they insisted, was still within the allowable
limits of "perfect working order":

Since I got the power supply back, I have run a 24 hour gel with standards
in 0.5 X TBE that  came out OK.  The normal gel that I am running,  with
10 times less EDTA hence, 0.5 X TBe, has resulted in a continuous smear.  
I also ran a PFGE gel on another machine, in another lab, with THE SAME
buffer fromthe THE SAME bottle, and it came out well.

I hope you will know something to tell me.  I have related this problem to
people all over the country, within my field and none of them have any
experience with this excpet to re-make their buffer.
Thank you, in advance, for your help. [email-masked]