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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”

Here’s the list (updated 5th May 2022 by JPKB): New updated schedule: Jamie Matt Isabel Caitie Delisa Jacquelyn Grace Roisin Jack Evan

View: bin 1, 0.2 s exposure Record: bin 1, 5s exposure Here’s how I set up SerialEM data collection on the Krios LMM.st  Save navi Align to marker  Go to 205x mag Close out of montage New montage – MMM Add squares Then check the MMM acquire script Acquire at items – MMM Find constrast-yContinue reading “SerialEM settings”

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”

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)”