Long Ranger caution. Answer my own question

Carla Drebing Carla.Drebing at UCHSC.edu
Mon Jul 1 09:10:08 EST 1996


	I posted a message this week about out of phase A/C and G/T peaks on my 
ABI 373 sequencer. My Amersham rep pointed me in the 
direction of one of their scientists who easily diagnosed the problem.  
The A and C are fluorescein and the G and T are rhodamine dyes. There is 
a two charge difference between the two and the differential migration of 
these is compensated for by the mobility files.  Running a new matrix 
file does nothing to repair this problem.  The Long Ranger gel recipe I 
was using ( double checked before I ever tried to use the stuff by 
confirming the method over the phone with FMC) does not analyze correctly 
with the 6% or the 4% primer mobility files.  Different mobility files 
screwed the data up in different places.  Alignment was better early to 
mid with the 6%, better later with the 4%.   FMC then told me they now 
have better parameters, %Ranger, TEMED, APS etc. for the 373 and are 
faxing the protocol.
	I had started using the Long Ranger with terminators and have not 
noticed as severe a problem, but caution to all who are using new gel 
chemistries.  Amersham's new primer system is coming with it's own 
mobility files, thanks.....  Thanks to Jim Tomfohrde of Amersham for 
finding the solution to a month of torture, when ABI, I assume unaware, 
had no idea it was the Long Ranger.

Carla Drebing
Univ of CO Health Sciences Center
Denver, CO 80262
303-399-8020 ex2722
drebingc at essex.hsc.colorado.edu




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