These past few months have been packed, so I haven't had a chance to write another post until now. I thought I should post an update before I graduate soon. Here's an update on my research and post-graduation plans.
Research
We have done a lot since my last post. With our previous methods, we figured out that the primers we were attempting to use for sequencing were not working. During our school's winter break, our team designed new primers and ran agarose gels with those. We finally got good results, and we were able to send in samples to Plasmidsaurus for linear amplicon sequencing. Something we didn't know with this new sequencing service was how the service did the sequencing. The samples we sent in were double stranded DNA, so when Plasmidsaurus inserted the sample into a plasmid, it didn't transform the way we intended and the data didn't turn out.
Next time we send in samples to Plasmidsaurus, we want them to be single-stranded, or to only isolate one allele. To accomplish this, we used the Restriction Enzyme Xclm. The initiation site for this enzyme to cut the DNA is within the region that CRISPR cuts with our gRNA. Our experiments consist of running a restriction digest on samples and running them on a gel along with samples that weren't digested, a normal WT, and a WT that went through the digest. Electrophoresis of these samples can show us if the DNA was edited by CRISPR or not. If the bands looked different from the WT that went through the digest, we could infer that it was edited. We then take these edited bands and cut them out of the gel. Using a gel extraction kit, we purify the DNA from the gel and prepare it for sequencing. Currently, we are troubleshooting how to get a sample that is concentrated enough. Our samples have been around 5ng/uL and we want 30ng/uL.
The goal is to send in samples by the end of the semester because 3 of the 4 current team members are graduating. The newer members are currently being trained, but we want to leave them with as much progress as possible. In the meantime, UWEC just had its annual research conference, CERCA (Celebration of Excellence in Research and Creative Activity). I presented with my research partner, and it was a fun experience to talk about my research with students at the same university as me.
Post-graduation Plans
In my last post, I talked about applying for post-baccalaureate programs. I ended up applying to 4 programs: Brown, John Hopkins, UW-Madison, and UMich-Ann Arbor. Two of the schools, Brown and John Hopkins, were reach schools, so I wasn't too surprised when I got rejected from their programs. I got to interview with UW-Madison back in March, but I also got rejected from their program. Lastly, I had an interview with UMich-Ann Arbor a few weeks ago, and I haven't heard back from them yet.
With the applications not going how I wanted, I started applying for jobs. I've been on multiple websites applying to lab technicians, research assistants, and other similar positions in the Wisconsin and Minnesota area. So far, I've gotten one reply from Exact Sciences. They are a Madison-based company that does genetic testing, cancer screenings, and much more. I interviewed for a position in their Rare Disease Extraction lab, where I would be preparing blood samples for DNA sequencing. The first interview went really well, and I have a second interview in a few days!
Although it hasn't gone how I planned, I'm actually excited about my future for the first time. I can see myself working in a lab for a few years before trying to reapply for graduate school, and I can see myself being happy about it. The rejections stung a little bit, but I believe that when one door closes another opens. After being in the school environment for around 18 years (with 5 being in college), it might be a good idea to step away from school for a bit. Either way, I will update everyone in a few weeks with confirmed post-graduation plans!