Mar 16, 2023
Viral purification from faecal sample
- sarah.schulz 1,2
- 1EMBL;
- 2NFDI4Microbiota
- NFDI4Microbiota
Protocol Citation: sarah.schulz 2023. Viral purification from faecal sample. protocols.io https://dx.doi.org/10.17504/protocols.io.81wgbywbovpk/v1
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: March 15, 2023
Last Modified: March 21, 2023
Protocol Integer ID: 78833
Keywords: viral purification from faecal sample protocol, purification of viral particle, viral purification, viral particle, faecal sample protocol, faecal sample, purification
Abstract
Protocol for the purification of viral particles from a faecal sample
Materials
SM Buffer, 0.2 μm syringe filter, 100-kDa-molecular-mass Amicon Ultra-15 Centrifugal Filter, DNase I , RNase
Troubleshooting
Homogenise 200 mg of faecal sample in 30 ml of SM buffer (50 mM Tris-HCl pH 7.5, 100 mM NaCl, 8 mM MgSO4)
Centrifuge sample at 4000 rpm for 20 min, then filter through a 0.2 μm filter
Add filtrate to a 100-kDa-molecular-mass Amicon Ultra-15 Centrifugal Filter
Centrifuge filter tube at 4000 rpm for 3 min to concentrate sample
Repeat centrifugation until the entire diluted filtrate has been passed through the filter tube
Dilute filtrate with 30 ml SM buffer
Add diluted filtrate to a fresh 100-kDa-molecular-mass Amicon Ultra-15 Centrifugal Filter
Repeat centrifugation until a filtrate volume of at least 500 μl is reached
Add 10 U of DNase I and 10 U of RNase to the filtrate and incubate at 37°C for 30 min
Incubate samples at 65°C for 15 min to inactivate enzymes
Store at 4°C