Today in the lab we made recombinant DNA. We cut two different kinds of plasmids - plasmids that encodes resistance to two different antibiotics - with restriction enzymes and then use ligase to combine them. I was not very prudent in following the procedures and made quite a few mistakes.
I made the first mistake when I was preparing the DNA samples for gel electrophoresis. I did not check the book for every step and I skipped the step in which we transfer some of the digested DNA into a new tube. I added loading dye directly to the digested DNA without transferring some of it to a new tube. I realized this mistake right after I made it, so I transferred the digested DNA, with some loading dye, to a new tube and then added some more loading dye to the new tube to ensure the loading dye concentration is as indicated in the book. However, the digesting DNA solution was not supposed to have loading dye in it and there was no way to take out loading dye from the solution. I made two more mistakes in this lab and the mistakes were probably caused by lack of sleep; therefore, I am going to sleep now instead of explaining the other two mistakes I made.