The summer of ChIP

Hi,

I’ve had a busy summer. All summer I have been working on my Chromatin Immunoprecipitation (ChIP) assay with the hope of showing that the Huntington protein can bind to DNA. After my second try I think I got it to work. However, the results were inconclusive since the no antibody smaple had the same fragment amplification as the sample with htt. Therefore, I think the antibody I used was not working, so I ordered a new anti-htt antibody which will hopefully allow me to successfully complete the assay. If I’m lucky by next week I should have results!

In addition to my ChIP, I have been working on growth assays and microscopy. To do this I needed to make new huntington plasmid constructions. I made a version of htt that glows red and cannot be SUMOylated, I will use this to see if htt localization is changed when there is no SUMOylation possible. Additionally, we made a htt that is on the activation domain (AD) of the GAL4 transcription factor. This will allow me to force htt to bind to the DNA and I can then see if it affects gene transcription downstream of the GAL4 promotor with ONPG and two hybrid assays.

I’ll keep you guys posted