Does anybody know a good method for large scale bacteriophage purification?

I am working with the coliphage MS2, the propagation protocol calls for chloroform as a lysis agent --> 20mL chloroform to 500mL culture. The chloroform is added for 15-20 min to lyse the cells thus releasing the virus particles to the medium. I then want to concentrate the virus via ultrafiltration. Prior to ultrafiltration I would remove the cell debris by centrifugation, 8000 RPM/ 15 min. I have read that chloroform can be removed by centrifugation. So will my centrifugation step to remove cell debris also get rid of the chloroform? Any other ideas to remove the chloroform? Or shouldn't I worry since I after ultrafiltration will replace the medium containing the chloroform with a Tris-buffer?

There is a method using PEG concentration, but I do not want to use chloroform to remove it because it may affect my Bacterial reduction assay.

How to measure the density of bacteriophages from a soil suspension like we measure the density of bacterial cells? I would like to know the phage density in the soil suspension to decide the inoculate density. Thanks in advance.

Would the dominance of one of the microbes in the gut change if we give different foods to insects or larvae? Suppose the insect is supplemented with amylase-producing bacteria, fed with carbohydrates such as rice, corn, and other sources.

Since humic acids share similar size and chemical characteristics to DNA, it is hard to purify DNA from it. It binds to DNA, fluorescence dyes and inhibits taq polymerase. I would appreciate any suggestions to get rid of it. I need the DNA for shotgun metagenome sequencing...

I have found information about using the kit (with modified protocols) for isolation of genomic DNA from filamentous fungi/cell culture/some bacteria. Has anyone tried this out on soil or sediment? I am not familiar with using Qiagen genomic tips at all, and so somewhat unsure of the requirements.

We are searching for any academician voluntary work to edit our journal for English proofreading. We are from the least developing countries and we can't afford to pay for this job but we are searching for voluntaries who are native English speakers.

I'm isolating bacteriophages from the faeces sample, but I can't see clear zones. I observe bacterial growth within the phage zones on the double agar plate that I left overnight.

Hi everyone,

I am trying to improve sustainability in my institute and I realized that there is an overuse of MilliQ water. I believe that for most of the operations done in a microbiology lab (buffers and bacterial media preparation, PCRs, restriction reactions, ligations, DNA assemblies, ...), using distilled water might be sufficient. I was wondering if someone has an overview of which water can be used for which protocol and maybe a list of the operations which really require MilliQ water.

Thank you very much in advance for your help!

Best,

Filippo