What are next steps after local refinement?

3D Refinement
1

Hi Everyone,

I finished 2D Classification, ab-initio, hetero- Homogeneous NU-.
I saw my molecules has two distinct domains A and B, and liker region is very flexible.
So I am testing local refinement for both domains.
But my knowledge is disconnected after local refinement.
Is there anyone can introduce further steps until getting final map?
For eg. How can I combine maps from two local refined map?
What cryosparc steps need for finalize?
Thanks

Jinseo

2

Hi Jinseo,

To combine two maps from local refinement, I would suggest using Chimera, taking the maximum value from the two maps at each voxel (command vop maximum) after alignment to a consensus refinement.

I would also suggest testing different post-processing approaches for your maps (CisTEM vs phenix.resolve_cryo_em vs relion, for example) and see which gives the most interpretable results.

What other steps are you not sure about?

Cheers
Oli

3

Hi Oli,

Is there any post refinements step are needed after combining map?
Thanks for your advice.

Jinseo

4

No - not if the individual local refinement maps have been postprocessed beforehand.

5

Thank you very much!