Crunch time
Hey guys! I’m sorry I haven’t been updating very often in the past few weeks, but it’s really been crunch time in lab. With only two more weeks left until I leave for the summer, I haven’t even gotten through ~half of what I had wanted to do. For example, I just finished my first whole mount in situ last night (Saturday) at 9pm. Now, I’ll have to section through the stained embryos in order to see the staining of the trigeminal ganglion. This in situ is the first of approx 5-10.
For those of you unfamiliar with in situ hybridization, it’s basically a staining protocol used to locate the expression of a particular RNA sequence; I use a DIG(digoxigenin)-labeled RNA probe that is designed to be complementary to my target sequence in order to do this (designing the probe actually takes quite a while as it consists of PCR, bacterial cloning/transformations, etc.). Once the RNA probe binds with the target sequence, an anti-DIG antibody is added to the tissue that causes a color change wherever the RNA probe has binded. Since I’m dealing with fairly young chick embryos, I am able to do a whole mount in situ, where the entire embryo is stained first and then sectioned through.
It’s been a busy couple of weeks and the next couple will continue to be just as packed with work. Honestly, 8 weeks is much shorter than I had expected!