Hello Dear Drs.
During the purification of bacteriophage stock by chloroform method prior to centrifugation and filtration .
Chloroform is able to lyse bacterial host cells . That means the DNA of bacterial host conc will be increased in the phage stock which will affect the purity of bacteriophage prepared for phage DNA extraction step .
is it better to centrifuge the bacteriophage stock without adding chloroform or adding chloroform is a must? And why ?
Hello, Chloroform was added to remove bacterial remnants, and efficiency of use is proved by many publications. If you want to receive the more pure fraction of bacteriophages so it must.
You can try using a filter sterilization step to remove bacterial cells yet let the smaller phages pass. this would let you get a higher purity of phage DNA at the cost of a lower yield.
Robert Adolf Brinzer
you are right
i thought the yield of bacteriophages will be lower because there are some bacteriophages are still inside host cells about to burst.
and The chloroform releases it .
Thanks for your answer Dr. Robert
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