Re: [ccp4bb] Unidentified density

+++1 Glycerol -- Kelvin Lau Protein production and structure core facility - PTPSP EPFL SV PTECH PTPSP AI 2146 (Bâtiment AI) Station 19 CH-1015 Lausanne Switzerland Email: kelvin@epfl.ch Phone: +41 21 69 34494 On 21 Feb 2023, at 14:34, David Lawson (JIC) mailto:davi

Re: [ccp4bb] TEV vs HRV3C

I agree. I think it depends on the lab and vectors and personal preference. But David, have you used them sequentially? I have once tried to make a His-GST-ENLYFQ-3C construct, and I found that it was self cleaving during expression. However I have never tried to replicate the results in vitro.

Re: [ccp4bb] CCP4i under M1Pro

go away when the Python3 version is ready to deploy. Best wishes, Randy Read > On 28 Jun 2022, at 16:45, Lau Kelvin > <5aaf8435dbef-dmarc-requ...@jiscmail.ac.uk<mailto:5aaf8435dbef-dmarc-requ...@jiscmail.ac.uk>> > wrote: > > Hello > > I have noticed ru

[ccp4bb] CCP4i under M1Pro

Hello I have noticed running normal programs like Phaser MR on a MacBook Pro (M1Pro) that jobs are extremely slow compared to my older computers. It doesn’t seem to be running over multiple CPUs anymore. Has this also been seen by others? Best Kelvin -- Kelvin Lau Protein production and str

Re: [ccp4bb] Eiger images and XDS

Hi Patrick I also just downloaded a Neggia library yesterday with data from SLS, on the newest XDS, and it was ok. Try the library here: https://github.com/dectris/neggia/releases However to me it looks like it cannot read the header information. Is the master.h5 file in the same directory as

Re: [ccp4bb] Glutaraldehyde and gel filtration

Ive always quenched and loaded directly. Never had a problem (for methylation reactions that are done in aldehydes) . However, you should ensure the column is already cleaned before and I would advise you to clean it after as well with NaOH, just in case you have excess glutaraldehyde and prote

[ccp4bb] Coot 0.9.4.1 EL and middle button centering

Hello I just updated to Coot 0.9.4.1 EL running on Big Sur and noticed that my middle button for centering does not seem to be working. All other mouse functions work. The previous version was just fine still. Did anyone else notice this? Best Kelvin -- Kelvin Lau https://people.epfl.ch/kel

Re: [ccp4bb] Micro/Macro crystal seeding experience

Wholeheartedly agree on all these points! Works wonders. -- Kelvin Lau https://people.epfl.ch/kelvin.lau Protein production and structure core facility - PTPSP EPFL SV PTECH PTPSP AI 2146 (Bâtiment AI) Station 19 CH-1015 Lausanne Switzerland Email: kelvin@epfl.ch Ph

Re: [ccp4bb] Looking for suggestions with protein expression

Hello Umar, I would not pin down your difficulties solely due to an Fe-S proteins. I have produced some with no fusion partners and they work wonderfully. They were expressed in an aerobic environment and then reduced in an anaerobic one before usage in reactions. 1) On the Fe-S side, there ar

Re: [ccp4bb] Accessing full list of programs in CCP4I2

: For occasions like Laus, it would be useful to further on have access to the CCP4 v7.0 Program Documentation, even if the programs are not updated as Christian explained. I was for instance looking for translating coordinates as in pdbset, but couldn't find a replacement in the ccp4i2 gui.

Re: [ccp4bb] Accessing full list of programs in CCP4I2

old fft task was discontinued. i2 works with coefficients, which generates smaller files and Coot provides all the functions to generate the map and is its own gui. Hope that explains a bit why things are how they are now. Cheers Christian On Tue, Jul 7, 2020 at 1:56 PM Lau Kelvin mailto:kelv

Re: [ccp4bb] Accessing full list of programs in CCP4I2

mport data, Molecular Replacemnt etc.) each point can be open up like a file tree to see all programs or pipelines available. You can search with the search field (Filter) on top for specific program names. Does that help? Cheers Christian On Mon, Jul 6, 2020 at 1:15 PM Lau Kelvin mailto:kelv

[ccp4bb] Accessing full list of programs in CCP4I2

Hello, I am looking for a way to find the list of programs accessible using the new 7.1 CCP4I2 interface? Is this still possible or do I have to revert back to version 7.0? Best regards, Kelvin -- Kelvin Lau https://people.epfl.ch/kelvin.lau Protein production and structure core facility - P

Re: [ccp4bb] Expi 293 (Met-) expression medium for selenomethionine labeling

e same, in my experience. But this was ~2 years ago so maybe Thermo made improvements in their production lines (I bet they did, this is a nice product and it ought to be a good money-maker for them) Artem - Cosmic Cats approve of this message On Sun, Mar 15, 2020 at 5:27 PM Lau Kelvin mailto:

Re: [ccp4bb] Expi 293 (Met-) expression medium for selenomethionine labeling

Dear Artem, By work poorly do you mean, not as well and are you referring to all Expi cells? We have adapted ExpiCHO cells to another medium and although we see drastic decrease from using the complete Expi system (x15 volumetric yield), it was still considerably higher than our previous cells