Cloning frustration

Sometimes the brilliance of my own stupidity explodes like a star over my head. I have been working on a cloning project off and on for a couple of months now in between other projects. About two weeks ago, I decided that I needed to get it done so that we could move on and finish the experiments that we had planned for these genes. The dumb cloning procedure is relatively easy but I just couldn't get anything to work. Then, a couple of days ago, I realized that the insert I am trying to add was single stranded DNA instead of the necessary double stranded DNA. I had neglected to order the complementary strand of DNA and therefore hadn't annealed the two strands together. What a stupid mistake. You'd think I was an undergrad doing this for my first time. I reordered my sequences of interest (a nuclear export sequence and a nuclear localization sequence) yesterday as double stranded DNA. Hopefully this experiement will work now.

I am also in the middle of treated my mice that we implanted. One of them has died but since the daily injections started by everybody else seems healthy. Only 10 more weeks to go.