Viral RNA extraction from Infected Tissues - Bertin Instruments

    Microbiology

    Viral RNA extraction from Infected Tissues

    Sources: at Leuven University

    Context

    In the context of one of the laboratories at the University of Leuven (Belgium), antiviral chemotherapy is studied and focused on. To evaluate the in vivo activity of the newly discovered antiviral compounds, we treated virus-infected mice with the antiviral compound, and extract the organs in which the virus replicates.

    Results

    After homogenizing the murine pancreas, Leuven University detects : – Firstly the viral RNA (1): The homogenization in lysis buffer RLT is performed, and worked further with the supernatant to extract the viral RNA. Graph (1) presents the evolution of viral RNA in the pancreas of viral-induced mice. It is expressed as “copies viral RNA per copy beta actin” (RT-QPCR after total RNA extraction). – Secondly replicating the infectious virus (2): In the second try (2) the homogenization in growth medium, (MEM, Gibco), is performed using saline buffer. The supernatant is then used to titrate on a cell culture (96-well plates, and the titer of the virus is determined (expressed as CCID50).

     

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    Graph(2) presents the evolution of infectious virus: the titer of infectious virus is expressed as “-log CCID50/ml” where CCID50 stands for “Cell Culture Infective Dose 50%”, which is a measure to express the titer of a virus stock.

    Graph(2) presents the evolution of infectious virus: the titer of infectious virus is expressed as “-log CCID50/ml” where CCID50 stands for “Cell Culture Infective Dose 50%”, which is a measure to express the titer of a virus stock.

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