Virus centrifugation purification method.

asked: Which man has done virus purification, can provide a virus centrifugal purification of the detailed protocol? Come on, thank you first!
: Virus purification has many methods, can be concentrated by chemical and chemical methods, adsorption method, speeding centrifugal method and so on. I used to use PEG precipitation method, relatively simple.
1, using ultrasound or freezing to break the cells to release the virus;
2, slowly stirring and adding NaCl final concentration is 0.5mol/L, which helps the precipitation of the virus, and then add 10% PEG in equal amount;
3, 4 to spend the night or stay for a period of time;
4, 8000/min centrifugation for 30 minutes to collect virus precipitation;
5, to add the virus precipitation to the appropriate amount of pbs, 4 to spend the night;
6, 10000/min centrifugation for 1 hour.
can also be further purified by gradient centrifugation or other methods.
: Your method is too helpful for me now, thank you. But how many places are not very clear, you can ask for advice code?
1, mine is chicken embryo bird cyst, the virus effect price is 1:640, only about 6 ml, how effective?
does it mean to add the equivalent 10% PEG in step 2, step two? What is the molecular weight of the PEG you use? Is 10% the concentration of PEG solution?
or collect precipitation after the last step of centrifugation?
: The general use of PEG6000 on it. If the purity of the sample is not strictly required, it can be recommended to use Millpore's
centrifugal enrichment
column, the Amicon series can concentrate 15 ml of samples at a time. Remember there is a document about PEG method precipitation HAV, you can go to download to see the reference. 10% PEG is, should be the final concentration. After centrifugation is precipitation, that is, concentrated virus.
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