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On Jun 10, 2009, at 9:23 AM, CW wrote:
You mentioned on tuesday you had a list of projects I can work on. Please send it to me if you have time (or bring it to class), I'm interested in working on something! So far, the mouse antibody project sounds exciting.
Date: Wed, Jun 10, 2009 at 2:44 PM
Subject: Re: Projects "shopping list"
1. I've got Taq polymerase in a plasmid in E. coli, Sha will probably do a crude harvest of it and we'll see if we can use this enzyme for PCR.
2. I've got a Peb1 gene in a plasmid in E.coli. I cloned this gene out of Campylobacter and some believe it is an important protein that helps Campy damage humans. The next work is to get E. coli to express the gene (feed them a chemical), break the cells and see if indeed the Peb1 protein is there. This involves running a protein gel and doing immunoblotting (Western). If Peb1 is being made we can then purify a lot of it and give it to JB... he'll inject it into mice and ultimately make antibodies against Peb1.
3. I have a black market way to build cloning plasmids, this a project I have to work on myself though.
4. We have some reagents to do Real-time PCR:
-harvest RNA from cells
-reverse transcribe the RNA into DNA
-use a PCR machine that has an optical head that measure the amount of DNA at each cycle of PCR (hence "Real-time")
-the goal would be to measure RNA levels (gene expression) of a gene between different cells (mouse v. human, cancer v. normal...)
Besides the DNA digestion work CM is doing the above are my more pressing projects. However, if you can think of something that is interesting, we could use in classes and are able to do out here with the given resources I'm all ears. Think about whatevers and we'll chat more whenevers.