I'm digesting a 11 kbp plasmid, resulting in three fragments (should be around 5, 4 and 2 kbp, respectively). What amount of digested DNA (in ng) would you recommend to be loaded onto a 1% agarose gel for subsequent gel extraction? What is the maximum?
Thanks for any helpful advice!
Carolin
Hi,
normally this info is written in the handbook. The Columns/ Beads have a binding capacity and therefore restrict the maximum amount to be isolaten. I would be rather careful from the gel perspective, as too much DNA might affect the separation efficiency. I normally digested and loaded 1-4µg and this always worked well for my cloning workflows
Sven
Sven Reister
I'm using the QIAquick Gel Extraction Kit, which should be working for up to 10 µg, but I guess that's too much for a gel. I will try with 1-4µg then! Thanks for your help!
Hi,
yes that's the way I would go (anyhow 2µg of Backbone is quite a bunch so should be more than enough)
Good luck
Sven
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