IUBio

Help with difficult template

radomski radomski
Mon May 5 08:36:37 EST 1997


I have been having trouble sequencing a plasmid's insert with 
dye-terminator chemistry. After about 20 bases into the insert, 
the signal drops to zero. 

The insert is 200bp and is composed of four 25bp repeats. So, "the deck is
stacked"  --  potential for hairpins?

I have tried DMSO in the cycling.

Any tricks, suggestions?

Or should I just continue with: formamide, or glycerol, and/or 
modify cycling to eliminate the annealing step (as ABI tech. help 
suggested) ?  


Thank you.


Chris Radomski
TerraGen Diversity Inc.
Vancouver, BC
Canada

radomski at terragen.com
radomski at unixg.ubc.ca









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