G blobs in Dye Term sequencing

From: Juliane K Mills (Juliane.K.Mills@dupontpharma.com)
Date: Thu May 31 2001 - 14:31:39 EDT


Dear Sequencing Core Facilities-
Does anyone else have trouble with G blobs? We used to have T blobs, so we switched from isoprop ppt to Edge columns for post-rxn clean-up before running on a 377 gel. That eliminated the problem (mostly) but now we're beset by these outrageous G blobs which appear around 50 bp and again between 120 bp and 130 bp. The interference gets worse the longer the cleaned, dried samples sit before being run (>36hrs). ABI has made suggestions about protecting samples from light and controlling humidity, but I'd appreciate any feedback or comments from anyone else experiencing this phenomena.

PS: We've seen this w/ BigDye ver2 and ver3.
-Juliane K. Mills
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Staff Scientist - Sequencing Facility
DuPont Pharmaceuticals Co.
E336/243E
office 302.695.2401
email Juliane.K.Mills@dupontpharma.com



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