No Band on agarose gel after Plasmid Isolation

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No Band on agarose gel after Plasmid Isolation

Post by MaraJade » Thu Oct 21, 2010 2:16 pm

Hi everyone,

when I make a plasmid isolation (Fermentas, GeneJet), I don't have any band on my gel. The Nanodrop says that I only have around 10 ng/ul. With other Plasmids it is working.

But I can make a transformation with this approach and I get some colonies on my agar plate, which grow on my antibiotic (kanamycin). When I make a PCR from this plasmid isolation I see some bands, but no expected bands.

The problem is that this is a mutated clone with a very good enzyme/gene and I must sequence it somehow.

Any suggestions?



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Inland Taipan
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Post by JackBean » Fri Oct 29, 2010 8:29 am

I've heard, that Nanodrop can tell you quite different values of concentration from the actuall. Maybe you have such a low concentration, that you cannot visualize it, yet there is something, so the transformation works ;) Are you able to do some PCR from that plasmid?

Cis or trans? That's what matters.

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