SpCase9-MBP fusion protein purification

The homologous Cas9 was fused downstream of the Maltose binding protein and a TEV protease cleave site. For bacterial expression of the recombinant proteins Cas9, corresponding plasmid was transformed into BL21-Plys (DE3)- competent cells (Stratagene). Expression of the recombinant proteins was induced by subculturing 5 ml starter culture into 1000 ml LB, adding at OD600Continue reading “SpCase9-MBP fusion protein purification”

Converting cryosparc to relion

1 – run cryosparc2star in the directory of interest, with the .cs particle file. Make sure to use the —relion2 flag.  csparc2star.py DrmABDNAcryosparc_P251_J73_008_particles.cs cryosparc-particles.star –relion2 2 – change .mrc to .mrcs within star file  sed -i ‘s/.mrc/.mrcs/g’ p205_j279_particles.star 3 – Create new extract job directory corresponding to the one in the cryosparc-particles.star file (use viContinue reading “Converting cryosparc to relion”

Introduce Yourself (Example Post)

This is an example post, originally published as part of Blogging University. Enroll in one of our ten programs, and start your blog right. You’re going to publish a post today. Don’t worry about how your blog looks. Don’t worry if you haven’t given it a name yet, or you’re feeling overwhelmed. Just click theContinue reading “Introduce Yourself (Example Post)”