1kb ladder kinase oddity.

Craig M. Story story at hydra.rose.brandeis.edu
Wed Sep 6 17:29:16 EST 1995


A few months ago I 32-P/kinase-labeled the Gibco 1kb ladder with fairly
good results on a sequencing (4%) gel. There were a few odd doublets
here and there where one band was expected. I repeated this recently
and now I see many extra distinct bands, particularly around the
506/517 area (about 12-13 monospaced bands clustered together), the 344
and 298 are also multiple bands. 

Has anyone else had similar experiences? 
Any theories on the source of the multiple bands? 

Note: the samples were heated well, and in formamide buffer, so I doubt
secondary structure, although the gel was not preheated...
 
Also, exonuclease may *not* be the problem, since there are single
bands there on the gel.

Finally, it is possible that what serves well on an agarose gel may not
be applicable for precise size standardization on sequencing gels. 

Craig M. Story

Rosenstiel Basic Medical Sciences Research Center
Brandeis University
Waltham, MA 02254

story at hydra.rose.brandeis.edu




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