I had an amazing last week in Boulder both in and out of the lab!
To start off the week I got to observe DNA extraction for the first time which was really cool! The Change Lab is currently waiting on more of their typical red blood cell lysis, so we were testing different brands of RBC lysis. As a result of the testing we had to use an RA’s blood instead of a participant’s, in case of any screw ups. The first step of DNA extraction is to use the RBC lysis to kill all of the red blood cells in the blood sample. After this is completed, cell lysis is added to kill of the remaining cells in the sample. The solution is hypotonic, so is floods the cells and eventually causes the cells to burst, thus killing them. Well both of the lysis solutions have been added and spun in the centrifuge, we poured the dead cells into a beaker of bleach. The cells clumped up in the beaker and created a kind of foam. It was really cool! After we let that sit for a while it got really warm to the touch which, according to the undergrad I was working with, means its killing off anything left over. Then the remaining parts in the sample are washed with a 70% ethanol solution to rid the DNA sample of any residual salts. IN the ethanol the DNA is visible, and it looks like little white strings! Then the final step is to turn the tubes upside down and let the remaining fluids drain out!
The next day I got to observe, and participate in, RNA extraction for the first time! It took us almost four and a half hours to complete the protocol! The blood samples we were extracting from were from 2015, but I learned that sometimes it just takes that long to get it done! The blood we were working with had been sitting in tubes with a cell lysis solution already in them, so all the samples were already lysed. The first step was to wipe everything down with RNAse away to get rid of any RNAses that might have been lying around in the bio cabinet. While we were cleaning the tubes were in the centrifuge. After they were done spinning I got to pour the samples out into a beaker until only the desired cell pallet at the bottom was left. Then we added nuclease free water(water that won’t bind to the DNA/RNA) and spun it again. After this spin we added the first of 5 buffers. I got to add the Br1 and use a pipette to re suspend the cell palette into the solution. The cabinet is really tiny, and you have to put your arms at a really weird angle to do this. After this we vortexed the tubes and then added the second buffer, Br2, and a protienase to get rid of any of the unnecessary proteins. Next the tubes are placed in an incubator for 10 minuets. For the next step we took tubes with purple column filters in them and added the solution to them. This acts as the third buffer. After being spun we threw away the columns and kept was was filtered through. Then we added ethanol to the remaining solution. After this we took pink column tubes and added the solution to them. For this stage we only kept what was in the filter and discarded what flowed through. Next, we added Br4 and spun the tubes, then repeated this step. Then we added a DNAse to the columns to kill any DNA that may have stuck to the RNA during the column spins. While we spun the tubes with the DNAse we also prepared a solution with a dye that binds to the RNA. Then I filled the tubes with the RNA sample with the dye. The final part of the RNA extraction protocol is called Qubit. Each tube is places in a Qubit machine(ours is painted to look like a lady bug) in which a light is shown that illuminates the dye stained RNA and quantifies it. This was one of the coolest wet lab protocols I have seen! I am so glad I was able to observe and take part in it!
For my last day at the lab my two main mentors bought pizza for everyone! There was a ton of pizza, so a lot of people from all departments of the lab stopped by and ate! It was so nice of them and I am so thankful I was able to learn so much while working with such wonderful people!
Outside the lab I had a great week! For the first part of the week the two older of my host siblings were out of town, so I got to hang out with just the younger one! We went the bounce house which is a building filled with all sorts of inflatable things to jump on. It was a lot of fun, especially for a four year old! Later in the week we drove down to visit a family friend of mine and her two awesome kids. They just put in a new pool, so the kids swam while I got to catch up with my friend! Then, on my last night in Boulder my host family and I rode our bikes to dinner and ice cream! The morning I left was quite an emotional one. My host family made me pancakes and bacon. All the kids wrote me cards and pictures which were adorable! They also gave me a super awesome t-shirt and a family photo for me to take!
I absolutely loved my experience in Boulder. I learned so much and had a great time working! I also had an amazing host family that I am going to miss greatly! I could not have asked for a better five weeks and I am so sad it is over!
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