Colony lifts - detection with biotinylated probes - background?

Peter M. Muriana muriana at aclcb.purdue.edu
Thu Sep 15 15:08:41 EST 1994


We are using biotinylated probes for detection of target sequences.  
Currently, we have started using them for detection of clones via
colony-lifts and have been experiencing background problems with the 
colonies.  Could this be due to cell-related biotin attracting the avidin-
alk.phos. conjugate?  Is there a way to get around this problem besides 
using dig-labelled probes and detection with anti-dig-alk.phos.?  I've 
noticed that Schleicher & Schuell's colony lift procedure with biotin-
labelled probes incorporates a step where they hand swab the lifts after 
they're lysed and baked, and then incubate them in proteinase K solution.
Just curious if others have experience such background problems, or not.
Regards, Peter
*************************************************************************
*  Peter M. Muriana, Ph.D.		Phone	= (317)-494-8284	*
*  Dept. of Food Science		FAX	= (317)-494-7953	*
*  Purdue University		E-mail	= muriana at aclcb.purdue.edu	*
*  W. Lafayette, IN  47907                                        	*
*************************************************************************
*	Disclaimer:  The opinions stated above are mine alone		* 
*	and do not represent endorsement by my employer			*



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