Hi,
I want to send my samples of phage for whole genome sequencing as we don't have the facility of WGS in our institute. I have extracted the DNA of phages by ZnCl2 precipitation method (Santos,1991). I'm not sure whether the DNA extracted by this method has enough concentration and purity which is required for WGS. Please give me suggestion whether this method is good enough or should i go for an alternative one?
I have not concentrated my phages by PEG or ultracentrifugation either.
Given that pfu/ml of my phage lysate are
- phage a= 1.3 × 1010
- phage b =2.1 × 108
I would start by measuring the DNA using a nanodrop. If you have access to a qubit or similar equipment, that would be ideal because the nanodrop may overestimate the concentration if the DNA is not pure. The amount of DNA needed would depend on the sequencing technology used (Illumina vs Ion Torrent for example) and how many samples will be included in the sequencing run. I would expect a couple of rounds to get a high phage titer.
@Tasha How much the concentration of phage DNA should be for sequencing purposes when we use nanodrop for measuring the DNA?
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