If it worked the first time, we would just call it search

I have been unsuccessful finding a clone that contains any one of my merlin constructs. It's a good thing that there is a plate full of bacterial colonies to pick through. I only need one colony to of each construct.

Today I am switching gears a bit and doing some cell culture. I have six plates of vestibular schwannoma cells that I am going to treat with a kinase inhibitor. I will let them incubate for a couple of hours, isolate the protein and run Western blots. Pretty straightforward and simple. I also have a couple of dissections to do this afternoon. I am going to remove the cochlea from a couple of mice. One of them is what is called a knockout mouse and the other is wild type. The knockout mouse has a gene, in this case a neurotrophin receptor, "knocked out" of it's genome. Additionally, one ear on each mouse has been deafened. I will begin by perfusion fixing the brain with paraformalydehyde. Then I will remove the temporal bone and proceed to dissect out the cochlea. After removing the stapes from the cochlea, I will make sure that both the round and oval windows are open and finally place the cochlea in fixative. Once fixed, I will de-calcify the bone with EDTA (yes, the same EDTA in the ingredient list on your box of Fruit Loops). Following de-calcification, we can section the entire cochlea into 7-10 µm thick slices, place them on a microscope slide, and label them with fluroescent tags to various proteins. By labeling the appropriate proteins, we can begin to get a sense of what role our neurotrophin receptor plays in deafness. I'll keep you updated at to our results.