FAQ  •  Register  •  Login

Benzonase and PSI clogging




Posts: 1

Joined: Wed Apr 27, 2022 12:06 am

Post Fri Apr 29, 2022 6:03 pm

Benzonase and PSI clogging

Hi everyone,

I am actually working on the optimization of a protocol for brain single cell suspension for CyTOF.

In the last few months I had to change 4 PSI due to clogging. The company thinks they are caused by the nature of my samples ( I had the same problem with PBMC) and for the next experiment they suggested me to try to use benzonase (https://www.sigmaaldrich.com/US/en/prod ... dlEALw_wcB)

I never used this reagent and I can't find any protocol about how to prepare it. Second, in which part of the staining should I use it? During the homogenization part or during the staining part?

Thank you in advance for your help :)



Posts: 4300

Joined: Fri Nov 01, 2013 5:30 pm

Location: Stanford HIMC, CA, USA

Post Tue May 03, 2022 3:16 pm

Re: Benzonase and PSI clogging

Hi Elisa,

Benzonase is a nuclease. It's added to media to help chew up any DNA released from dead or dying cells (cell processing, freeze-thaw, etc).

We add it at 1/10,000 to complete RPMI+FBS+Pen-strep when we're thawing cryoPBMCs. So, 1uL/10mL of media. I usually warm my media first, then add it just before I pull the cells out of the LN2. I then thaw each sample and add it to the Benz-media, spin down, resuspend in more Benz-media, and spin down again before counting cells and going on with my experiment.

Are you homogenizing and then going directly into staining, or are you homogenizing and then freezing?


Return to CyTOF general discussion

Who is online

Users browsing this forum: No registered users and 2 guests