Hi CryoSPARC users! We are excited to share with you a new end-to-end data processing case study of an active GPCR with a peptide ligand bound that you can follow along with!
Here we guide you all the way from downloading an EMPIAR dataset through to improving the quality of the particle stack, and separating different ligand poses, generating maps ready for model-building. Our final sharpened maps, half maps and masks are downloadable to allow comparison with your results.
This case study is of a membrane protein target at 140 kDa, and within the complete processing pipeline it includes ideas on how you might approach:
- Junk regions in micrographs
- Contamination of a particle stack with low quality or damaged particles
- Apparent late frame high frequency data in RBMC dose weighting
- Over-fitting in local refinement
- Separation and assessment of different peptide ligand binding poses
We hope this detailed guide is a useful resource for users old and new! If there are other types of targets, or particular single particle cryo-EM challenges that you would like to see a case study on in the future, let us know!
