Re: [ccp4bb] mosflm and pilatus 2M

2012-03-13 Thread Graeme Winter
Mosflm works fine with our Pilatus 2M at DLS - however you do need to be using the beta-test version. http://www.mrc-lmb.cam.ac.uk/harry/mosflm/betas/ best wishes, Graeme On 12 March 2012 19:57, Dean Derbyshire dean.derbysh...@medivir.com wrote: Hi again, it's the 2M  detector I'm having

Re: [ccp4bb] Envelope Phasing.

2012-03-13 Thread Petr Leiman
Using an envelope or a mask to start phasing even with high NCS is still worse than using a low res model from cryoEM that is characterized by an (almost) proper density distribution. One has to measure the intensities of the very low res reflections (0 0 1 and the likes) precisely. The

[ccp4bb] merge dataset

2012-03-13 Thread dengzq1987
Hi CCP4bb, I have two dataset from one crystal with different distance.now i am wondering how to merge the two dataset into one file,and use it to refinement. thanks a lot! deng

Re: [ccp4bb] merge dataset

2012-03-13 Thread Graeme Winter
Hello Deng, Process them as usual with e.g. Mosflm, then sort them together (you may need to rebatch one of the runs) and do one Scala run - this will put all of the measurement on a common scale and write out data suitable for refinement. For pointgroups with ambiguous origin choices (e.g. P4)

[ccp4bb] MR and pseudo-translationnal-symmetry

2012-03-13 Thread vincent Chaptal
Dear ccp4, I have a case of PTS and wonder what's the best strategy to handle my data. I processed my data in C2 with a=161 b=109 c=225 beta=104. The data in 97% complete to 3,8A. xtriage detected a 40% peak in the patterson at fractional coordinates x=-0,001 y=0,055 z=0,5. I want to try to

Re: [ccp4bb] MR and pseudo-translationnal-symmetry

2012-03-13 Thread Randy Read
Dear Vincent, The easiest thing for you to do right now in the CCP4 context is to run Molrep, with the flag for PTS turned on. If that works and gives a clear solution, that's great. If not, then I'd suggest downloading a recent version of Phenix and trying the new version of Phaser that

Re: [ccp4bb] MR and pseudo-translationnal-symmetry

2012-03-13 Thread Garib N Murshudov
There are 3 options: 1) Use molrep. If you have two copies related with PST then it can give right solution 2) Take the latest version of phaser and use it. It can deal with PST with two related copies also 3) Reduce cell dimensions (it does not seem to be possible in your case. PST is not

Re: [ccp4bb] Help! weird thing

2012-03-13 Thread Eleanor Dodson
That difference density looks too good to be true.. There is obviously a very strong non-crystallographic translation. Is the spacegroup and/or cell correct? Eleanor On Mar 11 2012, xiaoyazi2008 wrote: Hi All, I have an interesting thing to share. 2.3A dataset with good quality, P21

Re: [ccp4bb] Matthews coeff. from model

2012-03-13 Thread Eleanor Dodson
rwcontents xyzin my.pdb Eleanor It will not be able to guess the number og H atoms associated with the RNA so the answer will be a bit out. Eleanor On Mar 12 2012, james09 pruza wrote: Dear CCP4bbers, Is there any tool to calculate the Matthews coefficient from a crystallographic model

Re: [ccp4bb] MR and pseudo-translationnal-symmetry

2012-03-13 Thread Eleanor Dodson
I would try molrep - it will detect the NC translation and use it.. Eleanor On Mar 13 2012, vincent Chaptal wrote: Dear ccp4, I have a case of PTS and wonder what's the best strategy to handle my data. I processed my data in C2 with a=161 b=109 c=225 beta=104. The data in 97% complete to

[ccp4bb] Postdoc position available at the University of Oregon

2012-03-13 Thread Brad Nolen
The Nolen lab in the Institute of Molecular Biology at the University of Oregon is seeking a highly motivated postdoctoral researcher to join our lab full time on a flexible start date. Our interest is in understanding the molecular basis for the regulation of the cytoskeleton. We are currently

Re: [ccp4bb] My protein precipitates at r.t and dissolves at 4 oC

2012-03-13 Thread Kevin Jin
Hi Min, Please try this way if you use your protein for crystallization. 1. collect the needle and run SDS page or FPLC to verify the presence of protein. Make sure it is not a buffer salt. 2. You don't need to do dialysis to remove b-ME, otherwise it will take too long and you may lose some

Re: [ccp4bb] My protein precipitates at r.t and dissolves at 4 oC

2012-03-13 Thread Kevin Jin
Min, I forgot one more thing. If you try to grow crystall of membrane protein and get tiny crystal, you may try to add 5~10% IPA + 5mM NH4SO4 to addjuct the surface charge of your protein. The crystall may get larger. Folks like this trick. Good luck. Kevin On Tue, Mar 13, 2012 at 10:38

[ccp4bb] endotoxin removal: Triton X-114 dissociates protein oligomers?

2012-03-13 Thread Jerry McCully
Dear ALL; Thanks a lot for all the instructive suggestions. As my first trial, I tried 0.1%Triton X-114 plus Ni-column binding buffer. However, the oligomer of my protein got dissociated in to monomers as indicated by gel-filtration. Does anyone know how to

[ccp4bb] mammalian expression vector

2012-03-13 Thread Jerry McCully
Dear ALL; As an alternative strategy to avoid endotoxin, I plan to express the protein in mammalian cells. As suggested by others, the typical vector is pcDNA3.1(+). Does anyone have comments on this vector or recommend some other powerful vectors? I am new to mammalian