PFGE trouble>Hi there, > My problem with my PFGE apparatus is 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. 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. 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 from the THE SAME bottle, and it came out well. Thank you, in advance, for your help. nkh@sage.purdue.edu

Dunkles group dunkles_group at BTNY.purdue.edu
Fri Jan 26 10:37:54 EST 1996


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. nkh at sage.purdue.edu



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