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[DESCRIPTION]
Protocol for resuspending viruses from soils and sediments. See attached file for considerations, adaptations, and references. This version does not include CsCl purification, see version 2 for CsCl information. 
 

[BEFORE_START]
      Preparations
AKC Buffer (Make before and store at 4°C)
“Amended K-citrate”: 1% K-citrate + 10% PBS + 5 mM EDTA (don’t add if doing DNase) + 150 mM MgSO4; (per Liter: 10g of k-citrate, 1.44g of Na2HPO4 *7H2O, and 0.24g of KH2PO4 brought to pH 7)
1% BSA (w/v) in PBS
Need 2 ml per Amicon filter. Prepare 1% BSA (w/v) in PBS.
DNase
Weigh out sample in 50 ml centrifuge tubes

[GUIDELINES]
Need to record how much buffer is collected for viral counts
During the manual shaking, make sure the sample is broken up
The more new tubes the samples touch, the greater the viral loss will be
For the Amicon step, try not to use new filters. The filters will slowly get clogged and it may take a while depending on how many contaminants (e.g. organics) there are. Using new filters will increase viral loss. The same concept applies to the 0.2 µM filters. The composition of the filter may affect viral recovery.
For the DNA extraction, use the Qiagen PowerSoil DNA extraction kit
Need to do Swift library prep. If you want quantitative samples that have both ssDNA and dsDNA viruses
I have outlined areas where you can stop (store in 4°C overnight)
This protocol does not include EDTA in the AKC buffer. EDTA interferes with DNase and it mainly used to chelate metals. Don’t add unless you have high metal contamination
You will get contamination from any past or present viruses cultured or used in the lab

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