I have loved every moment of of my research project this summer. Although its been challenging adjusting to new terminology, vocabulary, and other hands on task, I could not ask for a better opportunity. My days in the lab have been pretty consistent for the most part, with few variations from the norm.
First I arrive around 10:30 where I go straight to my notebook to make sure I have documented everything I have done from previous days. Once I have done this I have a talk with my mentor, Ci, to understand what my next steps are in the project. Once we have had our discussion and I am sure that I understand what we are doing and the reasons why I begin setting up and preparing to run the experiment (most days this would be a PCR overlap). I write in my notebook every reagent that I will be using in order to ensure that I do not make any mistakes. Once I have completed this task, I begin conducting the experiment. I transfer the correct amounts of each primer and DNA template based on my master mix into PCR tubes.
Once I am sure that the correct reagents are in each tube I then place these tubes into a thermocycler (also known as a PCR machine). I program the machine based on the protocol of the polymerase that I am using. Once I confirm the time and temperature of each step, I start the machine to allow it to do its task of amplifying and constructing the DNA strands.
After the machine has finished amplifying the DNA strands, I take each PCR product and prepare it for gel electrophoresis. I do this by adding 5 micro-liters of 10x dye to each tube. Once this is finished I transfer each reagent into a separate well in the gel and run the gel in the electrophoresis machine.
Hopefully after doing this process I have successfully constructed and amplified the DNA strands that I need to continue my project. To confirm whether or not I have done this, I take a look at the gel using Ultraviolet light and see whether the position of the bands compared to the ladder matches my predicted outcome. If not, then I have to go back to the drawing board to see what went wrong. If so, then I will purify each PCR product that came out successfully and then continue on to the next step in the project. And that’s my day!
