molecular and radioactivity

Michael Sullivan via methods%40net.bio.net (by mlsulliv from wisc.edu)
Fri Dec 11 16:58:39 EST 2009


Although I don't use radioactivity often, there are still a few  
procedures where I do or would choose to use it.

For blotting/hybridization procedures, I would still choose  
radioactivity, hands down. As far as I know, most non-radioactive  
detection procedures require an immunoblot-like detection step. In my  
opinion, this just adds one more additional set of steps that needs  
to be optimized and opportunity for noise or background to be  
introduced inot the experiment. When might one want to do some type  
of hybridization experiment? A conventional library screen (I've done  
one of these in the last 2 years), outhern bolt to determination of  
the number of loci for a transgenic event, or looking at whether  
there are multiple classes of transcripts corresponding to a given  
gene. For the latter, if one wants quantitative data, radioactivity  
is linear over several orders of magnitude while most non-radioactive  
detection systems have a far more limited linear range.

Of course there are many biochemistry experiments that are most  
easily done with radioactivity, but this will be largely dependent on  
exactly what you are working on. I may be facing doing kinetics on a  
methyl transferase soon. From my perusal of the literature, this  
looks like it will be much simpler to do with radioactivity than by  
any other approach.

Mike



On Dec 10, 2009, at 5:43 PM, Hachey, John wrote:

>
> If you were to start up a molecular biology lab today, would you  
> require
> a setup for radioactive work? With all the various chromogenic,  
> chemiluminescent and fluorogenic
> detection reagents available today, is radioactivity extraneous?  
> Any specific protocols
> still absolutely requiring radioactivity?
>
> Thanks, Curious John
>
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