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Our lab investigates how the diversity of innate immune system genes impacts immune system function. Specifically, we study how polymorphisms of NK cell receptors called killer-cell immunoglobulin-like receptors (KIR) alter their NK cell function. We use an insect cell-expression system to produce recombinant proteins for use in a cell-free binding assay. To produce these proteins, we use a baculovirus transfection system in which our target gene is combined with linearized baculovirus and transfected into Sf9 insect cells. For this protocol to be successful, exactly 2ug of target gene DNA (in an appropriate display vector) must be added to 0.2ug linearized baculovirus. Given that we are often transfecting up to 20 samples at a time, establishing the concentration of target gene DNA in a timely fashion is of importance to us. The NanoDrop 8000 provides this utility for us – we use it primarily for the determination of nucleic acid concentrations – the fact that is has 8 channels that allow simultaneous measurement of 8 samples facilitates this process without compromising the accuracy of the measurement made.
The NanoDrop 8000 should be calibrated every 6 months (or more often depending on frequency of use) to ensure that the measurements made are accurate.
Product Name: NanoDrop 8000 SpectrophotometerSupplier Page