ECL for colony hybridization

dpetersn at unixg.ubc.ca dpetersn at unixg.ubc.ca
Mon Oct 24 22:09:38 EST 1994


In article <MURIANAP.11.00096EA4 at FOODSCI.PURDUE.EDU> MURIANAP at FOODSCI.PURDUE.EDU (Pete Muriana) writes:
>From: MURIANAP at FOODSCI.PURDUE.EDU (Pete Muriana)
>Subject: Re: ECL for colony hybridization
>Date: Sat, 22 Oct 1994 09:25:53

>In article <Pine.SUN.3.90.941021142929.9254A-100000 at chip.ucdavis.edu> Rinkei Ko
><ez034381 at chip.ucdavis.edu> writes:

I, too, had such problems, until I realized that I was no longer using nylon 
membranes.  I had gone back to the normal nitrocellulose circle filters I 
had used previously (Before DIG came to be) without realizing that the 
chemiluminescent reactants are repelled by nitrocellulose.  Check your 
manual to get a better description of this phenomena

>>I routinly use ECL 
non-radioactive DNA labeling system (Amersham) for >>Southern. Recently, I 
used same system for colony hybridization in order to >>clone a gene of 
interest. Instead of getting specific signals for the colony >>which has right 
plasmid, I got signals on all the colonies (False-positive).>>I wonder if this 
system is not suitable for colony hybridization because of>>non-specific 
protein-protein interaction, or washing condition was not >>stringent enough 
to avoid non-specific bindings.>>Rinkei Ko: rko at chip.ucdavis.edu

>Rinkei,



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