I'd use NCS in the beginning and maybe also DM solvent flattening,
then build and fix your structure, run refmac without NCS, superimpose
your chains in Coot and re-define your strict NCS groups accordingly.
Therafter you can submit your current coordinates to Ethan's TLSMD
server and continue from there
Jürgen
......................
Jürgen Bosch
Johns Hopkins Bloomberg School of Public Health
Department of Biochemistry & Molecular Biology
Johns Hopkins Malaria Research Institute
615 North Wolfe Street, W8708
Baltimore, MD 21205
Phone: +1-410-614-4742
Lab: +1-410-614-4894
Fax: +1-410-955-3655
http://web.mac.com/bosch_lab/
On Mar 28, 2010, at 18:34, Daniel Bonsor <[email protected]> wrote:
Hello again...
I have a 2.7A resolution data set. Spacegroup P212121 as suggested
by Pointless and best space group when run through Phaser. Unit cell
is 110.02x160.49x186.55. Mathew's suggest 11 complexes in ASU. Both
Molrep and Phaser can only find 6. This seems to be a typical
observation with the complexes that we solve (high solvent content).
Due to the low(ish) resolution I wish to apply TLS and NCS during
refinement. My question(s) is when to apply TLS and NCS, the order
in which it should be done, and when to switch off NCS (if at all)
during the stages of refinement. I have used a homologue complex
(Protein A -100% identical, Protein B - 80% identical) as a model.
I am open to any and all suggestions on the matter.
Thanks in advance
Dan
