question regarding making SS DNA

Bob Kodrzycki rjkodrz at westvaco.com
Tue Nov 19 08:49:43 EST 1996


Ahmet Arman wrote:
> 
> Hi netters,
> 
> In order to make functional assay, I have to isolate single stranded DNAs
>......
> my question is:1- Is there any good procedure for isolation of SS DNAs?
>                2-what is half-life of biotinylated primers?
> 
> Thak you for advance.
> Have you considered using a plasmid with an F1 filamentous phage origin 
of replication such as pUC119 (and several others ?Bluescript? I can't 
remember)?  Simply subclone your DNA of interest into the appropriate 
plasmid, grow up a liquid culture, and then infect with a helper phage.  
Single stranded DNA is produced selectively from one strand or the other 
depending on the plasmid used and secreted into the culture medium.  Just 
pellet the bacteria and isolate DNA from the remaining liquid medium.  
There is more info on this procedure in the DNA Cloning manual by 
Maniatis et al.  Or email me directly and I'll dig out more specifics if 
you are interested.

Bob Kodrzycki
rjkodrz at westvaco.com



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