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.
[cid:[email protected]]
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]> On Behalf Of Randy John Read
Sent: Monday, June 23, 2025 10:29 AM
To: [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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-----
Randy J. Read
Department of Haematology, University of Cambridge
Cambridge Institute for Medical Research Tel: +44 1223 336500
The Keith Peters Building
Hills Road E-mail: [email protected]<mailto:[email protected]>
Cambridge CB2 0XY, U.K. www-structmed.cimr.cam.ac.uk


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