week 6

We spent the 6 week performing mostly dot blots in order to determine the correct concentration of probe to use in the Pla assay. We also spent part of week trouble shooting because we were getting mixed results from our dot blots when using different concentrations of enzyme. We ran two Dot Blots in order to test if our concentrations were correct and to see if washing the nitrocellulose membrane in the Dot Blot apparatus had an effect on fluorescence. After developing our membrane we determine that washing the membrane in the Dot Blot apparatus was decreasing the intensity of the fluorescence. Once we knew that our dilutions were correct, we needed to come up with an explanation as to why we were seeing different results for our Dot Blots and Western Blot when using the same concentration of enzyme. This problem will be tested next week, and we will also get started on our Pla assays.