I am having bacterial contamination in my phages. Even after filtering my phage at 0.22um after a few months, there is a pellet formation occurring below. What should I do to remove contamination of my phage with bacteria?
Filtering with 0.22um filter should remove bacteria. It sounds like maybe you introduced the contamination after the filtration, perhaps by sampling with non sterile pipets or similar. For many phages you can store the phage stock with a few drops of chloroform to keep bacteria from growing, so you might see if your phage can tolerate chloroform and try that.
You can use chloroform to decontaminate those bacterial debris and after treating give a 5minutes spin at 10000rpm. Collect the supernatant.
Centrifuge and double filter sterilise sterilise. If phages concentration ist higher than E12 you will have a milky like suspenion. In this case you need to silute your stock. When phage precipitate ( like precipati g with PEG) they have grey color. In contrary bacterial precipate is white or other color depending on bacteria.
You can use chloroform followed by centerifugation and filtering with 0.22um
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