Mia Salas, Week 5+1 day of 6, Real-Time in Lab

This past week I gave a lab presentation at our meeting.  Kate and I both presented separately, and we each had to make a power point about our research.  I picked up donuts from Wawa on the walk over and passed them around before my presentation.  It went great, and my PI was very happy with the work I did over the summer.  I presented on NPC, starting with a basic outline of the disease and the goal of research moving forward, and then I displayed multiple excel graphs comparing the results when treatment began when the cats were 8 weeks old, 12 weeks old, and 16 weeks old.  I compared weight gain, a liver enzyme called ALT, and bilirubin, which is released when red blood cells are broken down, and then is carried to the liver, made into bile, and excreted out of the body.  A main conclusion that I and the people from my lab derived from my research was that there is not a substantial difference between cats who started treatment at 8 weeks old versus 12 weeks old.    

Towards the end of the week, Dr. Vite sent us to work in another lab with Ping and her daughter Gloria to get some hands on lab experience.  I sort of wish we were working with Gloria the whole time, because she is so great and she lets us do our own PCR, enzyme assays, and so much more.  She just graduated from UPenn, and now she works with her mom, studying epilepsy and MPS.  It was such a change from observing and working with tons of records and excel to putting on a lab coat, gloves, and helping a grad student with her research.  She loves teaching, and she explains every step.  Today we had to do DNA extraction from blood and PCR real-time to determine the genotype of each of the dogs: affected, heterozygous, or normal.  PCR-real time is a machine that quickly allows us to view the genotype of the dogs after about an hour to an hour and a half of putting the PCR samples in the machine.  PCR real-time is different from standard PCR in that in addition to adding two primers, we have to add in DNA probe in order to actually see the progress of the amplification on the computer.  The DNA probe is fluorescently labeled, and so with each cycle of PCR, the fluorescence increases.  As PCR occurs, a graph begins to form on the computer displaying cycles of PCR on the x axis and fluorescence on the y axis, and eventually the graph takes on an exponential shape.  Gloria said that real-time PCR is quicker than standard PCR and running the gel afterward, we can analyze the data easier with quantitative values, and we can actually see if the reactions in PCR are working as they happen.  Gloria will show us tomorrow the final results of this PCR.  We also looked at the results of our DNA sequencing from the week before; she uses DNA star, which is basically the same thing as DNA subway that we used in biotech.  We tried to look for any differences in nucleotides, but the only 2 that we were excited about turned out to commonly differ between dogs, and had nothing to do with epilepsy.  I also went to a lab meeting with Gloria and the rest of the lab.  This lab was mainly younger grad students who were eager to explain everything to me and Kate.  The rest of this week I’ll be working with Gloria!

Other than the lab, some friends and I spent the night in Philly going out and sleeping over.  It was fun to really explore the city and see what it was like at night.  In the morning we went for a run, so I finally got in some miles for XC (basically the extent of my summer training haha).  But running in Philly was so much nicer than at home, and Derek knew a 4 mile loop that was so perfect for running. Also, I must seem like a really approachable person, because the amount of times I get stopped with questions about trains, the Lucy, buses, or just in general how to get places is unreal.  But I don’t really mind; it’s cool being able to answer all of them.  I can’t believe this will already be my last week in lab.