DNA sequencing problem

Dale Levitzke Dlevitzke at medicine.newcastle.edu.au
Mon Feb 12 20:19:03 EST 1996


It seems that not all of your excess dye terminators are being removed 
from your reactions before you load them so try the following.

1.  If using 3M Sodium Acetate make sure it is at the correct pH.
2.  When removing your ethanol and acetate use a pippete tip to make 
sure all of the excess ethanol/acetate is removed from your pellet.
3.  Do a 70% wash and make sure the pellet is dry before you add your 
formamide loading buffer, and once again make sure all the excess 
ethanol is removed.

I have been using the ethanol protocol for some time and it generally 
gives excellent results, good luck.





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