Thank you Prof Raaij. I also have bad experience with sonication for one proteins. It denatures after lysis with sonicator (1sec on/off pulse, 30% amp, 5 min). However, when we lyse the cell with beads (vortexing with beads for 1 min, 5 times), we were able to get good protein.
Best Regards Veerendra Kumar, PhD Associate Professor & Ramalingaswami Fellow Amity Institute of Molecular Medicine & Stem Cell Research (AIMMSCR) J-3 108, Amity University Campus Sector-125, Noida-201303 UP, INDIA http://amity.edu/aimmscr/Faculty.asp Google Scholar: https://scholar.google.com/citations?user=ymdnsUAAAAAJ&hl=en From: Mark J. van Raaij <[email protected]> Sent: 23 March 2021 18:13 To: Dr. Veerendra Kumar <[email protected]> Subject: Re: [ccp4bb] E.coli cell lysis using glass beads In my opinion it is better than sonication, but more work, especially if you want to wash the beads and recycle them. So I haven’t been able to convince people in the lab to use it more than a few times. French press could still be better or about the same, not sure. I also think it should affect protein quality less than sonication, perhaps about the same as French press. Mark J van Raaij Dpto de Estructura de Macromoleculas Centro Nacional de Biotecnologia - CSIC calle Darwin 3 E-28049 Madrid, Spain Section Editor Acta Crystallographica F https://journals.iucr.org/f/ On 23 Mar 2021, at 13:23, Dr. Veerendra Kumar <[email protected]<mailto:[email protected]>> wrote: Dear All, Sorry for the off topic. May I ask your experience about bacterial cell lysis using glass beads beating? How is the cell lysis compared with sonication and French press? Does the glass beads affect the protein quality? Thank you in advance. Best Regards Veerendra Kumar, PhD Associate Professor & Ramalingaswami Fellow Amity Institute of Molecular Medicine & Stem Cell Research (AIMMSCR) J-3 108, Amity University Campus Sector-125, Noida-201303 UP, INDIA http://amity.edu/aimmscr/Faculty.asp Google Scholar: https://scholar.google.com/citations?user=ymdnsUAAAAAJ&hl=en ________________________________ To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/WA-JISC.exe?SUBED1=CCP4BB&A=1 ######################################################################## To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/WA-JISC.exe?SUBED1=CCP4BB&A=1 This message was issued to members of www.jiscmail.ac.uk/CCP4BB, a mailing list hosted by www.jiscmail.ac.uk, terms & conditions are available at https://www.jiscmail.ac.uk/policyandsecurity/
