Livah Method Calculations for qPCR Analysis

     This week we discussed the math behind our qPCR analysis, and learned the steps to do it ourselves. While we do get the calculations from a computer, its always good to be able to double check the calculations, and to be able to find where an error is if their is one. We covered the calculations for relative expression (called the livah mathod), not absolute expression as that's the calculations used in our project and also used for most qPCR calculations. 

    After covering how to do the calculations, we also discussed and reviewed our procedure. We hadn't been getting no significant change in gene expression when under oxidative stress from H2O2. Even when tracking the KatA gene, which should have light up with H2O2, we saw no significant change. When looking at the paper we were basing our procedures on, we saw that they actually halted the H2O2 reaction by adding catalase. After a small discussion, we decided that next week when moving forward with our procedure we would be adding catalase after the H2O2 reaction to end it. Hopefully this will give us better results and show us some gene expression. 


Below are my notes from how to calculate relative gene expression-






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