SYBR Gold Cloning

Bas Jansen bas at nospam.nl
Tue Jan 25 17:03:15 EST 2000


In article <v04210103b4b3b5b4300a@[134.121.168.66]>, chris at genomex.com 
(Chris Small) wrote:

> Hi,
> 
> I was wondering whether anyone has experience staining high MW DNA 
> with SYBR Gold, visualizing it with the "Dark Reader" then isolating 
> it from the gel and cloning it.  Some literature says it doesn't 
> damage the DNA (like ETBR) and subsequently affect cloning efficiency 
> but I would like hear it from someone that has tried it.
> 
> Thanks in advance!

Hello Chris,

I have used SYBR Green in combination with the DarkReader, when 
isolating cut'n cipped pBluescript and pGEM vectors. Cloned two 
fragments (also stained with SYBR Green) in both vectors, and judging by 
the amount of colonies it works just as good or better than EtBr 
stained, UV-exposed DNA. The big plus is that you don't have the hassle 
with EtBr and UV, and you get a nice pair of glasses with the DarkReader!

Best regards,
Bas

-- 
bjhj at dds.nl




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