True and the “more faulty” those reflections are the bigger is their influence. 
Would then it make sense to flag possibly “faulty” reflections not to be used 
in the free set? I think it is a logical conclusion, however, recognizing the 
faultiness may not be as straightforward, in my opinion.

Thank you for your comments.

Vaheh

From: Jeroen Mesters <[email protected]>
Sent: Monday, June 23, 2025 12:39 PM
To: Oganesyan, Vaheh <[email protected]>
Subject: Re: [ccp4bb] free R in shells

Hi,

with 5% of the reflections randomly flagged by numbers 0 to 19, any of these 
Rfree sets is 19x smaller than the working set of reflections. The impact of a 
few really faulty reflections in terms of Intensity will therefore be much 
larger on the Rfree set than on the Rwork.


Regards,

Jeroen

__
Dr. math. et dis. nat. Jeroen R. Mesters
Biological Safety Officer (BBS)
Deputy, Lecturer, Program Coordinator Infection Biology
Visiting Professorship in Biophysics - University South Bohemia

[cid:[email protected]]

University of Lübeck
Center for Structural and Cell Biology in Medicine
Institute of Biochemistry
Ratzeburger Allee 160
23562 Lübeck

Tel +49 451 3101 3105
https://orcid.org/0000-0001-8532-6699<https://orcid.org/0000-0001-8532-6699>


Am 23.06.2025 um 16:55 schrieb Oganesyan, Vaheh 
<[email protected]<mailto:[email protected]>>:

Hi all,

I’ve spent some time going through different flags for free reflections for 
(somewhat stupid) reason to get lower Rfree. I made sure all different flags 
were used only for each of sets of refinement. I wanted to satisfy my inner 
belief that it doesn’t matter which flag is used (between 0 and 19). To be 
short: I failed.

Free reflections have been chosen “randomly”, no suspicion there. However, 
there was clear difference at the end. The difference was about 2-3% difference 
in Rfree value. This tells me that the randomness has some sort of rule, which 
makes “random” choice not so random. Having found this I also tried to see “a 
rule” that breaks this randomness, like even numbers, odds, etc. I did not. 
Because this was only on few cases I won’t even try to connect it to # of 
molecules per AU, or SG. For each structure (I tried this for 2-3 structures 
about 10 years ago) it was a different flag.
I’m not sure if DCC also is looking through different flags, but at the end it 
finds the best, making these exercises unnecessary.
Sorry, cannot present a case. Was too long ago.

Vaheh Oganesyan, Ph.D.
<image001.png>
R&D | Biologics Engineering
One Medimmune Way, Gaithersburg, MD 20878
T:  301-398-5851
[email protected]<mailto:[email protected]>



From: CCP4 bulletin board <[email protected]<mailto:[email protected]>> 
On Behalf Of Randy John Read
Sent: Monday, June 23, 2025 10:29 AM
To: [email protected]<mailto:[email protected]>
Subject: Re: [ccp4bb] free R in shells

Hi Ben,

I would be very interested if you have a case where it makes a difference to do 
this. At one point I was convinced that it had been important when we were 
working on the structure of a Shiga-like toxin bound to trisaccharide (1bos), 
with four pentamers in the asymmetric unit. However, Pavel Afonine challenged 
me to show that the free set was less biased when chosen in shells than when 
chosen randomly, and even in that relatively extreme case I couldn’t see 
evidence of it. So it’s probably not worth the bother. Also, if you select the 
free set randomly, it’s distributed over the same resolutions as the working 
data, which arguably is important when you’re using it to calibrate the 
sigma(A) estimates for likelihood targets.

Best wishes,

Randy

> On 23 Jun 2025, at 12:35, Ben Bax 
> <[email protected]<mailto:[email protected]>> wrote:
>
> Hi,
> How do you select R-free in shells with CCP4?
> Thanks, Ben Bax
>
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