My experience at the Fred Hutch explorers program has been great, I have learned so much and I have gained tons of knowledge on the inner workings of lab work. Many of the labs in this program incorporate the fundamentals of lab work alongside the bits of information about biology. One of the labs I wanted to talk about today was a lab focused on testing for Chronic Myeloid Leukemia. The purpose of this lab was to simulate isolating sample DNA from patients to detect BCR-ABl gene using gel electrophoresis. Gel electrophoresis is a process used to separate DNA fragments, this is done so the strands can be placed into a well at one end of a gel that is a part of an electrical current. Inside that electrical current it is expected for DNA strands to be pulled towards that positive side of the gel, as DNA strands are negatively charged. The DNA attraction towards the positive side depends on what charge the DNA strand has, meaning the result for each gel electrophoresis can differ depending on which strands are being observed. In our case, we are trying to observe the BCR-ABL gene. By looking at the DNA, we can see which DNA is most likely to be inside of the gene based on the del electrophoresis. When going through the procedure, one of the most important things to do was to make sure our microliter values for each time we used the micropipette were correct. If they were off, we would end up with more or less of each substance. We first started by getting our patient sample card, which contained a blood sample from 4 patients. We first had to cut out the blood samples using a hole puncher and place them each into individual microtubes. The next thing we had to do was deposit 100 microliter of a Rehydration Solution, this solution is used to liquify the blood sample, this will create a patient sample, a mixture of the solution and the DNA of the patient.
After obtaining the patient sample, we added 50 microliters of each patient sample to a 96-well plate and sent it for pcr processing, which is used to detect genetic material.
In the next steps, our goal was to load our patient samples into a gel box to perform gel electrophoresis. Me and my lab partner had 8 columns on the gel plate, 4 of these were control samples, samples used to show the expected outcome in the gel depending on the circumstances of the DNA strand. These included a DNA ladder to showcase the DNA strands, two positive columns (one showcasing short and long strands) to show what would happen if a patient had a positive result, a negative column to show what would happen if a patient did not have anything, and the 4 patient columns. The goal was to evaluate how each of the 4 patient columns related to the control brackets, allowing us to determine whether the patient was positive or negative.
The next section focused on us micropipetting our samples into the brackets of the gel. This section was a bit difficult, as we both had to make sure that each of us pipetted the right amount of each substance to make sure we had the right amount needed to have solid data. We also had to make sure to not mix up any of the tubes, as there were a lot, so placing the substance of one tube in the wrong bracket was something to be careful of.
In our result, patients 4,7, and 8, all had negative results, however, patient 3 had a positive result. The strand showing patient 3 was positive was faint, but me and my partner could still see the short strand indicating this.
When comparing our results with the class, we found that other group members had similar results to what we had, meaning our experiment was successful.
This experiment was a very unique experience for me. It allowed me to use the knowledge we had on procedure and using it in an actual lab was really nice. It allowed me to get a feel of what a career resolving around this experiment would be like. I am really glad I participated in this lab, and I hope I will use the knowledge and experience gained from this lab in the future.