*NO* DNA eluted from plasmid prep - Why? - (Jul/11/2015 )

That makes sense. Thanks again.

I would suggest you always make sure that you are indeed using Kanamycin in your broth when making the plates and not ampicillin for example. You wouldn't believe just how often a simple mistake like that can happen when cloning. I would also try to do a colony PCR before growing your cells in the flask as you can test if the plasmid is there before doing clean-up etc.

Hi HawkeyeGrad,

I set up a control situation to test the effectiveness of the Kanamycin today.

I put my bacteria with plasmid in LB/Kan (made fresh), put ampicillin-resistant bacteria in the same media, and then plain media in the incubator to ensure what is supposed to grow is indeed growing, and that the Kanamycin is effective in preventing unwanted growth.

After some research, apparently Kanamycin has a short life in solution (read: five days)...so it is probable that this is causing the issue.

We shall see tomorrow what the Kanamycin is and is not doing!

Thanks again for all of your input.