So far I have had a lot of success obtaining highly concentrated DNA containing my Presenilin D275A mutant. In order to do this,I have produced and utilized competent cells to propagate the abundance of DNA. Then, through lysing the cells and “collecting” the DNA, I am able to obtain a high concentration of my target DNA that will be used to synthesize RNA.
One of the procedures I have become extremely efficient is the ability to demonstrate my data through gel electrophoresis. Gel electrophoresis is the used in the lab to separate DNA by size (length in base pairs) for visualization and/or purification. Electrophoresis uses an electrical field to move the negatively charged DNA toward a positive electrode through an agarose gel matrix. The gel allows shorter DNA fragments to migrate more quickly than larger ones. Thus, you can accurately determine the length of a DNA segment by running it on an agarose gel alongside a DNA ladder (a collection of DNA fragments of known lengths). Here is an example of the picture we got from running a gel:
I can’t wait to continue my research begin RNA synthesis soon! Updates to come!