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Loni Pickle, Life Technologies R&D Scientist, covers some key pointers for moving into tissue-based chromatin immunoprecipitation (ChIP) experiments. ...
Loni Pickle, Life Technologies R&D Scientist, covers some key pointers for moving into tissue-based chromatin immunoprecipitation (ChIP) experiments.
Hey fellow ChIP researchers, and anyone new to the ChIP application, I'm Loni Pickle, fellow bench scientist, here to talk to you today about optimizing your ChIP, giving you some tips that I've learn from spectacular mentors over the years, spending a lot of time at the bench myself optimizing ChIP and also helping you guys out there in the field. So I wanted to bring you what I've learned here today. So what we're gonna talk about is tissue ChIP; we're gonna go through magnetic beads and how they're gonna make your life much easier when performing ChIP, and we're also gonna talk about shearing and shearing optimization."
Tissue Processing Using Magnetic Beads Shearing Optimization
Let's start with tissue processing for ChIP; a very important step in your ChIP application. When I moved from cells to tissue I was looking for a protocol that I could use. What I found out there it took too long, required me to purchase more expensive equipment, and it required a majority of my sample; which I wanted to use that sample for other purposes. So what I did was, we kinda developed our own protocol. Basically using a gradient of syringe needles; which made it quicker, made it extremely cheap, and cut down the amount of material to much, much less; meaning I could use that material for other purposes.
The important things that will be needed for this procedure are:
- 1 ml syringes
- 18 and 21 gauge needles
- 1 X PBS
- Petri Dishes
- Standard ChIP Reagents (formaldehyde, Lysis Buffer, and Glycine)
The five basic steps we are going to go over today are:
- Homogenizing with gradient needles
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