I have made PR8 H1N1 influenza virus by reverse genetics system and then validated by HA assay. But when i infect the A549 and HEK293 cells with the same infection is not happening after 24 or 48 hrs. What could be the reason for that?
Ashish what is the HA titre of the virus you are taking for infection purpose? You can try infecting with the titre of 8 or 16 and try using MDCK cells instead. Also please check that the pH of the media you are keeping the cells in is alright.
I did it with those cells and it worked really well. Maybe you have to add TPCK-Trypsin to "activate" the HA.
Ashish,
How do you infect the cells? knowing that will help us to answer your question.
Good Luck
Hanan
Even i tried NS1 mutant of PR8 for the interferon induction. But couldn't see any induction of Type I interferon.
Dear Ivan Haralampiev, can you plz send me the exact protocol which you are using.
1-The HA of your rescued virus is low and HA is not an indicator for representing infectious virus. So, it is better do concentrate the virus by ultra -filtration.
2- Do blind passage for several times in order to adaptation of the rescued virus to new cell lines.
- Badges
- Science method