Double Stranded DNA Virome of the Bovine Rumen
The rumen content was centrifuged at low speed to separate larger particles and then filtered using 0.2u filters to separate out bacteria and other cells. Next, the supernatant was centrifuged with PEG8000 to co-precipitate the viral particles away from other small molecules. Then, a CsCl gradient centrifugation was performed to separate out the viral particles organelles such as mitochondria and chloroplasts. At this point we should be left with protein capsid-covered viral particles. Finally, the samples were treated with DNase to remove any non-protected DNA and followed with proteinase K treatment to release the viral DNA. This viral DNA was digested with restriction enzymes, ligated adapters, and PCR amplified.