Welcome to crystallography and the PhD. Expect a lot of failure before your degree. I know its frustrating but all of the challenges you encounter will teach you and increase your skills. I had crystals my first year, moderate resolution around 3 A and it took until end of my 4th year to phase a DNA binding domain bound to DNA and then another 1.5 years to refine, generate mutants and test binding.
Dealing with lab misinformation, although unfortunate and annoying, is also part of the process. You don't say what your resolution was. New crystals, protein or dna constructs or even additives might make a difference. Did you test a room temp crystal to make sure it's not your cryo effecting resolutions? Hang in there. A phd is like climbing a mountain or a marathon, one step at a time will eventually get you there even if it doesn't feel like you're making progress. Mary Mary Ho Postdoc Arnold Lab On Tue, Jul 13, 2010 at 5:36 PM, Xun Lu <xlun...@gmail.com> wrote: > Dear All, > > I took over this project last September. Since the information in the > lab are not well managed, it's kind of starting over (WRONG plasmids > sequences; even WRONG oligo length/sequence in the complex which has been > successfully crystallized. Those crystals only gave low resolution data.) My > boss told me to mutate the Cys to Ser, see if it would help. I finally got > many perfect-looking crystals, and sent them to synchrotron. We didn't get > better data at all. I think it makes sense, because the crystallization > condition is similar to the one they got previously (I have tried their > condition first of course, but at that time, the DNA oligoes were wrong!) > > I am trying MBP fusion now, but am very upset these days that I have > made some stupid mistakes. How to cheer up? And most importantly, what > should I do to improve the resolution? I heard a talk on a conference about > microbeam. AND, a student in another crystallography lab in the same > department has tried microbeam and got VERY GOOD data which far far beyond > her and all others' expectation. In our lab, we didn't even know they were > going to try the microbeam in chicago. Also, anyone can share experience > with dehydrating crystals? > > Although I didn't get good data this time, I have to say that, I have > learnt a LOT during crystal screening, cryo, and remote control of > synchrontron beamline from people in other labs. > > > Thanks, > > Xun > first year PhD > > >
