probing rRNA with biotin labelled oligos

Sharee sharee.mccammon at agsci.utas.edu.au
Thu Feb 9 22:33:46 EST 1995



Has anyone out there ever probed rRNA (16S, to be specific) on
nitrocellulose with biotin labelled oligos? It should have been quite
straight forward......at least that is what I used to think! I've managed
to piece together a protocol from published methods which used
radioactively labelled oligos and others using biotin labelled larger
probes - surprise, surprise, its not working. I'm getting  non-specific
binding and almost no specific binding. I'm currently playing around with
hyb and wash stringencies, but would be hugely grateful to anyone who care
to share their experience  with this protocol with a frustrated netter. 
Thanks in advance
Sharee
PS I'll even put the names of anyone who can help me on my Xmas card list!



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