SBB09Ntbk-Madhvi Venkatesh: Difference between revisions
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==[[User:Madhvi Venkatesh|Madhvi Venkatesh]] | ==[[User:Madhvi Venkatesh|Madhvi Venkatesh]] 15:24, 23 February 2009 (EST)== | ||
So I rewrote | So I rewrote my construction files after I realized that I can't display a protein on the N and C terminal ends of fliD because they are not facing the surface. The new versions of the construction files including Chris's notes on them are linked from the construction files page. We got oligos last Wednesday and I set up PCRs for the N and C terminal parts and wobble PCRs for the linkers. I purified and digested over the weekend. I also set up the SOEing PCR at the end of last week. Today I ligated the parts into pBca1256 and transformed. I will pick colonies tomorrow and send for sequencing on Wednesday. I also purified and digested the SOEing PCr for removing the restriction site in the N terminal fliD part. | ||
Here are gel pics from the PCRs: | |||
[[Image:MV_2-23-09.jpg|200px]] | |||
Revision as of 13:24, 23 February 2009
Madhvi Venkatesh 14:54, 2 February 2009 (EST)
Today, I got my project ball and I am making a new flagellar display system. I need to consider several possibilities and figure out what would be best to make and the source. The project description if on this page: new flagellar display system
Madhvi Venkatesh 15:40, 9 February 2009 (EST)
Possible places to find EtpA homolog (from CTEC):
APEC 01:k1 CFT073 EHEC 0157:H7 Shigella flexneri
Madhvi Venkatesh 15:24, 23 February 2009 (EST)
So I rewrote my construction files after I realized that I can't display a protein on the N and C terminal ends of fliD because they are not facing the surface. The new versions of the construction files including Chris's notes on them are linked from the construction files page. We got oligos last Wednesday and I set up PCRs for the N and C terminal parts and wobble PCRs for the linkers. I purified and digested over the weekend. I also set up the SOEing PCR at the end of last week. Today I ligated the parts into pBca1256 and transformed. I will pick colonies tomorrow and send for sequencing on Wednesday. I also purified and digested the SOEing PCr for removing the restriction site in the N terminal fliD part.
Here are gel pics from the PCRs:
