Thursday, April 7, 2011
ChIP update
Science moves at a snail's pace most times. I finally have the first part of this experiment nailed down (DNA isolation and cleavage). I was using a sonicator to break up the DNA but was getting poor results. I went to an enzymatic method called Micrococcal Nuclease digestion and have had some success. I am going to run through this once more tomorrow and attempt to clean the results up by combining the MNase digestion with sonication. Next week I will move on to infecting the cells with virus so that p75 will be expressed and then work on getting p75 binding to beads for the immunoprecipitation part of the experiment. Once I have that worked out, I will change cell types and make whatever small changes I need to make before really hitting this experiment for results.
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3 comments:
I've just started reading the book about Henrietta Lacks (HeLa cells). Very interesting read so far. Do you ever use human cells or just mice cells?
Jason, I knew you were nuts, but now Chelsey, too. This is bad.
I would like to know where do you learn to do all of this stuff. You didn't just learn this in a book.
Dad Clark
All the cells I use in culture are human cells. We don't use HeLa cells but they are the most used cell line in the world. As a matter of fact, it is estimated that the majority of cell lines are contaminated with HeLa cells.
Henrietta Lacks' story is an interesting one. Her cervical cells were really the start of modern cell biology.
Although I am currently using a cell line for the preliminary part of this experiment, I will be using primary human cells from a Vestibular Schwannoma to gather the results that we will use for future publications.
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