I detected several new viruses in my target insect through NGS and RT-PCR. Now I would like to use S2 cells to see if I can extract infectious viruses from these infected field insects. I read that we can do it through homogenizing bugs in chilled medium and passing the supernatant through 0.22 filter and infecting cells. Does anybody have the full protocol?
Hi,
The protocol depends on the size of your insect. But what you have described as the methodology is completely right. You would not be able to pass the homogenized insect solution through 0.22 um filter as it would block it right away. You have to pass the homogenate through a cheese cloth first and then a 0.45um filter to avoid cloging the filters. The last step would be 0.22 um filter. You can use normal medium or PBS to make your insect homogen solution. Let me know if you need more details
Thank you Leila joon:)
How about if I first centrifuge homogenized insects and then filter the supernatant instead of using a cheese cloth?
One more question: How do you infect cells with the filtered solution? Directly adding it to the cells or what?
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