We would like to separate the 30 and 10 kb digestion fragments of an NcoI digestion of a 40 kb linear DNA. They should not only be separated from each other, but also from undigested 40 kb DNA. Any preferred low-cost methods?
I'd think that they should be resolvable on a 0.5% agarose gel. Try using a high range ladder: http://www.thermoscientificbio.com/nucleic-acid-electrophoresis/generuler-high-range-dna-ladder-10171-to-48502-bp/
You should be able to purify your dna by gel extraction. Although, if you add sufficient enzyme, no ciruclar plasmid should be left uncut.
Actually, I would use 0.4% agarose gel and no more than 5V/cm... On the other hand, pulsed field gel electrophoresis would be my primary choice :)
Hi Mark,
maybe you have seen it already, but in Life Technologies have a Tips&Tricks page to help you with this:
http://www.lifetechnologies.com/es/en/home/life-science/pcr/elevate-pcr-research/agarose-content-with-tips-and-tricks.html#2
My experience tells me that a 0.5% agarose gel should be OK but...
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