Sep 01, 2020
Version 1
Ultracentrifuge clean-up of phage lysate for TEM V.1
This protocol is a draft, published without a DOI.
- 1University of California, San Francisco
Protocol Citation: Kathy N Lam 2020. Ultracentrifuge clean-up of phage lysate for TEM. protocols.io https://protocols.io/view/ultracentrifuge-clean-up-of-phage-lysate-for-tem-bkmkku4w
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: September 01, 2020
Last Modified: September 01, 2020
Protocol Integer ID: 41356
Take 1.1 ml of existing plate lysate
Take 1 ml to clean up; save remainder to titer pre- versus post-clean up
Transfer 1 ml to ultracentrifuge tube
Centrifuge 25,000 rpm (~25,000g) 60 min 4C
Pour off supernatant away from pellet; pipette off residual
Wash with 0.1 M ammonium acetate
Centrifuge 25,000 rpm (~25,000g) 60 min 4C
Pour off supernatant away from pellet; pipette off residual
Wash with 0.1 M ammonium acetate
Centrifuge 25,000 rpm (~25,000g) 60 min 4C
Pour off supernatant away from pellet; pipette off residual
Resuspend in 0.1 M ammonium acetate
Titer pre- and post-clean up samples
Store at 4C for up to 2 weeks