Thanks for the suggestions so far, I should have given a little more
information in my initial post. The protein I'm working on is an Gram-Neg
Beta-Barrel about 100kDa, likely with 22 strands.
I'm currently crystallising in bOG, although my next optimisation step is to
try a range of detergents
With information you've provided I have multiple suggestions for you, some
of which you may have already tried:
1. OMPs can be fairly particular about which detergents they will
crystallize in. Try exchanging the protein into a different
detergent/detergent mixture and then set up new trays in
Hi Rhys,
I suspect that what you call a gel might be phase separation (correct me if
this is wrong) like an oily phase enriched in protein and detergent. you
may have too much detergent in your drop.
may I ask what detergent you are using (low or high CMC) and at what
concentration of detergent do
Hello Joe,
You're either going to have to supply your script with something like a
.sca file and extract unit cell and space group from it, or you'll have
to supply those values on the command line.
First option in csh, apropos Jan's script:
#!/bin/csh -f
#
# run as "./phils_script1.csh dumm
Dear All,
I have 2 positions available for software developers in cryoEM:
1) A developer to work alongside Chris Wood on the CCP-EM project. The post
will support a number of software development projects (see e.g.
http://www.ccpem.ac.uk/ccpem_projects.php), as well as providing general
commun
Hi All,
A quick question if you've ever worked on membrane proteins, I'm trying to
optimize crystals for bacterial integral outer membrane protein I'm working on.
I'm getting some fairly modest rod like crystals in a 0.1M Tris pH 7.5, 30%
PEG 600, 0.03M MgCl2 condition. However I get lots and
You can always synthesize your own too!
Artem
Or idtdna. Dna2.0, blue sky, and many others
On May 9, 2013 10:01 AM, "Mathew Martin"
wrote:
> Hi Randy and Rodger,
>
> We have had real success using BlueSky (MA) for insect expression, I
> believe we have had around 15-20 genes synthesis-optimized
At this point you do have the scalepack2mtz output file (BTW,
imosflm/aimless is wholeheartedly recommended by this convert), and you
can easily extract all the info from there. There is mtzdmp, of course,
but it's easier to parse the actual mtz file (hey, the records are
actually text). Like so:
Hi Randy and Rodger,
We have had real success using BlueSky (MA) for insect expression, I believe we
have had around 15-20 genes synthesis-optimized with these guys. The expression
levels have been rather reasonable too (obviously protein dependent, but on
average 1 mg of xtal grad protein per
I hope this isn't too much of a thread hijack, but i also wanted to add a
question to Randy's : How big a protein can one express in insect cells?
I've read in a few places that it's not size limited but i wonder if
that's borne out in practice.
On 9 May 2013 16:05, Randy Watson wrote:
> Hi a
Hi all,
Sorry for the off-topic request. I am considering ordering codon-optimized
genes for expression in insect cells (High Five). I wonder if anyone could
make suggestions regarding: which companies are best to order from
(accurate synthesis – cost efficiency) and what optimized codon sequenc
Looks like my signature messed up the script too...
scalepack2mtz hklin test.sca hklout test.mtz << eof eof
^^^
Typo:
To avoid crash/overwrite, should be
scalepack2mtz hklin test.sca hklout test.mtz << eof eof
^^^
jbb
Jeffrey Bonanno, Ph.D.
New York Structural Genomics Research Consortium
-Original Message-
From: CCP4 bulletin board [
The deadline for articles submitted for the next issue of the CCP4 Newsletter
is Friday 14th June 2013.
Articles may cover any topic of interest to macromolecular crystallographers,
though we are particularly interested in articles on software or methodology.
Short items of news are also very w
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Dear Joe,
most ccp4 programs make use of space group and cell information if
present in the input file. The script you list below does not have an
input file (apparently it creates a dummy mtz-file with only a list of
Miller indices), so there is noth
Hi all,
Graeme's message tell the core of the detector usage notes here at diamond. He
might have unearthed a can of worms or two, though ;)
CCDs still are great detectors! I am not sure about his noise comment as the
more recent CCDs (post 2000-ish) are designed to operate with anode sources
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