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Problem with NIH 3T3 cells


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#1 Sravi

Sravi

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Posted 23 August 2011 - 12:46 PM

Hello everyone,

I am growing NIH 3T3 cells in lab and using them to infect with adenovirus. So every time i do infection i used to keep 4 wells as a control in 12 well plates. The problem is when i am doing the Beta-glo assay on both infected cells and control cells, the control cells itself giving very high RLUs. I used to get 20,000 RLUs previously. Now i am getting 150,000 RLUs.

Infection Procedure:

1. I am seeding the 3T3 cells in 3*10^5 ratio in 12 well plates the day before infection. and after 18 hours of seeding i am infecting those cells with adenovirus. For control cells i am not infecting. after 48 hours of infection i am doing beta glo assay on them.


Need help...Please pour your suggestions.




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