[ccp4bb] CCDs + Re: PILATUS data collection

2013-05-09 Thread Jose Brandao-Neto
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

Re: [ccp4bb] question about CCP4 scripts

2013-05-09 Thread Tim Gruene
-BEGIN PGP SIGNED MESSAGE- Hash: SHA1 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

[ccp4bb] Reminder - CCP4 Newsletter submissions

2013-05-09 Thread Karen McIntyre
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

Re: [ccp4bb] question about CCP4 scripts

2013-05-09 Thread Jeffrey B Bonanno
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

Re: [ccp4bb] question about CCP4 scripts

2013-05-09 Thread Jeffrey B Bonanno
Looks like my signature messed up the script too... scalepack2mtz hklin test.sca hklout test.mtz eof eof ^^^

[ccp4bb] Codon Optimization for Insect Expression

2013-05-09 Thread Randy Watson
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

Re: [ccp4bb] Codon Optimization for Insect Expression

2013-05-09 Thread Roger Pickman
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

Re: [ccp4bb] Codon Optimization for Insect Expression

2013-05-09 Thread Mathew Martin
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

Re: [ccp4bb] question about CCP4 scripts

2013-05-09 Thread Ed Pozharski
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

Re: [ccp4bb] Codon Optimization for Insect Expression

2013-05-09 Thread Artem Evdokimov
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 mathew.mar...@rocketmail.com wrote: Hi Randy and Rodger, We have had real success using BlueSky (MA) for insect expression, I believe we have had around 15-20

[ccp4bb] Membrane Protein Optimisation

2013-05-09 Thread RHYS GRINTER
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

[ccp4bb] 2 positions available for software developers in cryoEM

2013-05-09 Thread Martyn Winn
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

Re: [ccp4bb] Membrane Protein Optimisation

2013-05-09 Thread Pascal Egea
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

Re: [ccp4bb] Membrane Protein Optimisation

2013-05-09 Thread Jim Fairman
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

Re: [ccp4bb] Membrane Protein Optimisation

2013-05-09 Thread RHYS GRINTER
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