So, I'm in the process of moving to Sweden. A long, drawn-out, painful process. I don't have a job there yet, I haven't graduated from here yet and yet!
It was kind of nice to hear that they sent out application off to Sweden without needing an interview. Makes me feel like something is going the right way!!! On the other hand, I now have one less opportunity to take a forced vacation...D'oh!
I wonder if I can get the BF/fiance to get some sort of Indian residency visa, similar to the one that I will get for Sweden. Then he will have an equally easy time traveling to India as I will to Sweden.
Tuesday, September 13, 2011
Wednesday, August 25, 2010
15 is for "I really need a vacation."
Had my part IIs yesterday and got a lot of good ideas but mostly need to start working on the two projects that I have.
1) Streptavidin evolution with the unnatural
-For this I need to check if IDT will make me a DTB-primer.
-I want to see if some of matt's solubility mutants work in my system. If they do this will mean I might be able to skip the system evolving solubility mutants from my variant pool and focus instead on evolving the DTB-binding variants already!
I am looking for good ways to get a randomly mutated pool and I printed out Matsumura and Ellington's protocol for the same. I know there is also sexual PCR and the MnCl2 PCR, I think I need to go over those again. There is also what looks like a nice reference in it called, "The effect of high-frequency random mutagenesis on in vitro protein evolution: a study on TEM-1 beta lactamase." from J. Mol Bio in 1999. Looks pretty relevant. I think this is the most important task in this project going forward.
-Andy suggests getting some PURE system in here and testing to see how that works in comparison to my stuff. Doable. Lets see.
2)Rationally designed yeast synthetase
- M.Winkler suggested testing to see if glutathione works better than DTT. I need to check this the next time I make a reaction mix, maybe do a couple of reactions to check.
- I need to take out the LacIq portions from all the synthetase plasmids and try to make lysates with them.
- Need to make primers to PCR up the synthetase mutants.
Ok, thats it for today. I'm kinda done. And I need a vacation.
1) Streptavidin evolution with the unnatural
-For this I need to check if IDT will make me a DTB-primer.
-I want to see if some of matt's solubility mutants work in my system. If they do this will mean I might be able to skip the system evolving solubility mutants from my variant pool and focus instead on evolving the DTB-binding variants already!
I am looking for good ways to get a randomly mutated pool and I printed out Matsumura and Ellington's protocol for the same. I know there is also sexual PCR and the MnCl2 PCR, I think I need to go over those again. There is also what looks like a nice reference in it called, "The effect of high-frequency random mutagenesis on in vitro protein evolution: a study on TEM-1 beta lactamase." from J. Mol Bio in 1999. Looks pretty relevant. I think this is the most important task in this project going forward.
-Andy suggests getting some PURE system in here and testing to see how that works in comparison to my stuff. Doable. Lets see.
2)Rationally designed yeast synthetase
- M.Winkler suggested testing to see if glutathione works better than DTT. I need to check this the next time I make a reaction mix, maybe do a couple of reactions to check.
- I need to take out the LacIq portions from all the synthetase plasmids and try to make lysates with them.
- Need to make primers to PCR up the synthetase mutants.
Ok, thats it for today. I'm kinda done. And I need a vacation.
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