Can you sed a bit of your scalepack unmerged data - that would allow us
to check format and pointless behavior..
It sounds a bit like a scalepack problem though..
Eleanor
Alexandra Deaconescu wrote:
Dear all:
I am trying to solve a structure from apparently a hexagonal crystal.
I indexed
Dear all,
We are attempting to clone into pETDuet, pACYCDuet, pCola1Duet and pCDFDuet
and we are encountering numerous difficulties.
We are aware that most of them are low copy number, but even taking this
into account, we find it puzzling that whereas the first preps of the
vectors gave
This may be of interest to some of you.
We are in the process of putting together what we hope to be an exciting
program of talks with plenty of opportunity for discussion.
Liz
Dr. Liz Duke
Principal Beamline Scientist
Diamond Light Source
Harwell Science and Innovation Campus
Chilton
OX11 0DE
Hi,
This is not typical. Try different cell lines for propagation and also add
more sugar and glycerol to your medium - this helps.
Artem
Dear all,
We are attempting to clone into pETDuet, pACYCDuet, pCola1Duet and
pCDFDuet
and we are encountering numerous difficulties.
We are aware that
Dear BB!
I have a strange problem with TLS refinement and refmac.
In my AU there are 4 monomers of the same protein.
Based on structural subdomains I defined 3 TLS groups for each of those
monomers so that the overall TLS group number is 12.
When I do TLS and restrained refinement from the GUI
Dear Silvia,
we have cloned genes in pETDuet, pACYCDuet and pCDFDuet without any
problems.
What we always do, with any plasmid, is transform bacteria freshly
before any miniprep or protein preparation - we do not believe in
stocking plasmids on plate or as glycerol stocks. Glycerol stocks
I dont understand it either but I have also found there are problems
with starting again from the previous end point..
If you have run TLSANL after the refinement, then the TLSOUT would be
totally inappropriate to start from..
The TLS records n the PDB file and the ANISOU records are
Dear Eleanor,
Even if Denzo, Scalepack and HKL suite have been extensively developed,
their file formats have not changed in many years. However, there are
intermediate programs between Scalepack and structure solution ones that
can potentially be a source of problem. For example, ctruncate for
Hi Matthias,
a few suggestions:
- use TLSMD server to define TLS groups;
- before using TLSMD do some group ADP refinement with one or two
refinable ADP per residue (you can do it in phenix.refine), so you get
less biased ADPs from previous restrained B-factor refinement;
- just out of
Alex,
With regard to scalepack, be sure you use NO MERGE ORIGINAL INDEX, not
just NO MERGE.
Marian Szebenyi
MacCHESS
Dear all:
I am trying to solve a structure from apparently a hexagonal crystal.
I indexed and scaled data in P6 in Scalepack (with merging) then used
Scalepack2mtz (with
agree with Michael, use low concentration of detergent. I normally resuspend
the cell in buffer and sonicate, spin down at 6000 g., resuspend again with
buffer containing small amount of detergent and resonicate, spin down again
at 6000 g. finally was with buffer again (resuspend and spin down).
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