Colony Hybridisation with biotinylated oligo

Mr. A.F. Hill ahill at crc.ac.uk
Wed Jan 20 07:45:58 EST 1993


Hi

I'm trying to set up a system for doing colony hybridisations using a
biotinylated oligo (23-mer) and using streptavidin-biotin-alkaline phosphatase
and NBT/BCIP to get the colour reaction...

most protocols i have read are for using probes labelled via nick-translation
or random priming and im wondering whether an oligo with one biotin group
will work the same...

ive spotted the oligo onto a filter and can read a signal down to 10pg but
when i hybe with the oligo and develop the colour i get a nice purple
membrane...

can anyone help?

thanks...

Andy

==============================================================================
 Andy Hill				  |		
 Prion Disease Group			  |     E-Mail: ahill at uk.ac.crc	
 Dept. Biochemistry & Molecular Genetics  |-----------------------------------  
 St Mary's Hospital Medical School	  |     Phone: 071 723 1252 ext. 5469
 LONDON W2 1PG				  |     Fax:   071 706 3272
==============================================================================



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