So this happened.
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So this happened.

#hi jenny! hope you don’t mind if I reblog this #I know you’re going through some hard times but I find your singular self determination quite inspiring #I hope I can similarly pursue an area of interest with such commitment and passion #when I figure my shit out #the last line is the most valid thing I’ve heard about success. - elysiantears

Hey! Oh my gosh, I had so much trouble trying to read the hashtags when I realized I could just go on your blog to see it all. I was having such a shitty day. This made me really happy. Thank you :)

I think you will be amazing. I’m excited to see the way your future will unfold because you’re very unique. Good luck with the rest of high school :P and keep blogging because I read it all :D

(I’m posting this because I rarely get a compliment like this hahahaha.)

These past few weeks I’ve thoroughly immersed myself into the project that I am working on at the moment. Not only because I want to have my work published, but I want to be a primary author, or coauthor of the paper. 

My dad asked me why I was working so hard when I could go work with him and get my name on one of their papers quickly and easily. But I think there is such a huge difference between getting something through connections compared to getting something as a reward for hard work. I am lacking in things I can genuinely be proud of. The work my current boss does is not exactly close to the type of research I want to do in the future but I think it’s all so interesting and exciting so it’s not bad in any way. Also, I’m happy that I am able to work completely independently on my own project with the possibility of publishing everything I’m doing within a year’s time. I think this way, I gain a lot more through trial and error and just all the typical mistakes and hardships one would typically face in a lab setting. It’s really easy to memorize protocols and do stuff but it’s really hard to get things perfect enough for publishing. It’s hard to come up with original solutions when you’ve exhausted every banal quick fix. It’s even harder to conjure up innovative novel ideas. It’s why I’m at the lab from 9-6/7 and work right through weekends on a similar schedule. It’s also confirmed I will be working on my project through the school year as well which is exciting because I don’t really know how school is going to be for me and the lab is somewhere I feel completely comfortable.

It’s so tough. I mean I’ve done many westerns. Since grade 10, I’ve done westerns. You’d think it’s easy by now. And sure, it is, in some respects. I no longer forget things, I can make the gels blindfolded. But to get beautifully perfect bands… Oh my god. You think you’ve solved one problem, and another emerges. It’s crazy. You realize eventually: Everything must be perfect.

In addition, I’ve witnessed some really hard working people. I mean, there are a lot more people who don’t give a shit, but when you start working on weekends regularly, you begin to see who’s what kind of person. There are a few people I’ve seen that come in every weekend and always work super late as well. They make you open your eyes to the fact that you can definitely push yourself harder. They motivate you in some ways. You see what kind of person gets into U of T’s MD/PhD program. You become that person naturally.

I think it’s good for me. Especially now that I’m heading into university, seeing graduate students regretting their lives, makes me know that every moment matters. Every opportunity matters. And that sometimes you absolutely must make your own opportunities because they rarely come knocking at your front door. Time wasted is time you’ll never get back and sometimes you must feel vulnerable and hopeless and angry and frustrated to be able to learn. Passion is only fifty percent of the battle, the rest is hard work. I used to think that because I was so passionate about the things I wanted to do that I was guaranteed a place in this world to do it. But the uncertainty of it all is so frightening now, there is no choice but to work like it’s the last thing you’ll ever do.

So I woke up at 8:30 am this morning and felt the need to go back to the lab to finish up what I was doing on Saturday.

I get off the bus at Eglinton. I forget my access card. This would be okay if it was a weekday but it’s the weekend. So I bus back home. Grab my access card, go back.

I get there. Did some calculations. Started to set up my westerns. Realized we were out of running buffer. The recipe on the container was messed and pretty much I had no idea how to make running buffer/I googled but didn’t know where the stuff was. Long story short, I figured it out eventually, but I couldn’t find masks anywhere and inhaled an amount of SDS worth a lifetime. Great. Legit took me an hour to figure out.

Moving on. I prepared my samples from my lysis on Saturday. Set up six westerns gels. Started loading my samples and the sample decides to float out of the well. This normally doesn’t happen. 

So I did some troubleshooting. At first by myself.

My initial reaction was that my samples were too sticky because my boss doesn’t want me to centrifuge my samples. Normally you centrifuge your samples, because the stickiness is due to DNA and you can pellet that out by centrifugation. But I thought it was strange that my samples were still “sticky” because I upped my lysis buffer volume from 150uL to 230uL. The stickiness was resolved at 200uL but I still decided to up to volume just to be absolutely sure. So vortex my samples some more to mix. Still doesn’t work. I tried my other samples. They all floated out of the wells.

So I text my boss to see if I could run my samples through a syringe. See, you lose a lot by doing that, so I wasn’t sure if she wanted me to. She told me to add 50uL more of my sample buffer, reboil my samples, and that I shouldn’t do the syringe thing on my own. So I try to reboil my samples. (I only did 6 to see). The dry bath broke down right then and there. So then, I was on a mission to find another dry bath. After another good amount of time, it still didn’t work. 

At this point, there was another summer student at the lab so I asked him. He “supervised” me while I ran one of my samples through a syringe. Annoying as shit process. Still didn’t work.

His girlfriend suggested I add 1:1 water to my sample buffer to see if that was the issue. Even the water mixture floated out. Hinting that my sample buffer was somehow contaminated.  Then she suggested I reboil my samples and not put them on ice. Didn’t work. 

At this point, I call my dad because it was already the afternoon, I had already spent a majority of my day there fucking troubleshooting, I was not about to leave empty handed. He did a good amount of yelling at me with regards to precision when it comes to lab work and hung up on me eventually.

Feeling like all 18 of my samples were about to go down the garbage chute (12 of which were my siRNA knockdown plus the protein concentrations were perfect this time around), I burst into tears.

I look up feeling like there was too much liquid on my gloves. My nose was bleeding uncontrollably. So I bleed out for a good 30 minutes in the bathroom and at this point I wanted to go home so badly. I was just done with life. But I realized I had everything still out I had to clean up and my nose still wouldn’t stop bleeding. And then I realized one of my samples was still boiling, and it had already been close to 40 minutes, I’m panicked. I legit run around the lab, still bleeding uncontrollably, clean up with one hand, and run back to the bathroom to try and stop myself from bleeding. Thank god it was a Sunday, the women’s bathroom looked like someone died in the sink.

I go home. Depressed. Sleep. Wake up. Looked up the composition of the sample buffer. Glycerol is the stuff that makes the samples heavy enough to sink into the wells. Went back to the lab at 10:00 pm. Put 50uL of glycerol in my samples.

And it worked.

So fucking stupid. Whoever decided to contaminate the sample buffer and not say anything. 

(But while I was there, I found that two of my samples wouldn’t freeze meaning they must have been contaminated with ethanol or something of the sort.)

Please promise me that you will work really hard and be successful and happy and have everything that you’ve ever wanted. 

It makes me sick to know that you’re not happy with yourself right now and that there’s nothing I could possibly do to alleviate your frustrations. But you still have so much time and so much potential. Please, please promise me you will try your best, and in four years, when I talk to you again, you’ll be happy. Promise me that. 

In four years, I want to be in medical school. But I can’t be happy unless you are too. We’ll both work really hard, okay? In four years, I want to hear about all the amazing opportunities you’ve been presented with. I want to hear how excited you are. I want to hear you tell me how happy you are again. Promise me you will.

Sunday lab selfie 😁
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Sunday lab selfie 😁

You’re probably going to dance with another girl who will taste like fresh picked strawberries and smell like flowers blossom in her hair,

and you’re probably going to choke down 5 shots of straight vodka and get the thought of me out of your head and focus on the girl dancing with you who wants to be your apple pie but you can’t see the diamonds in her eyes because you’re staring at the ones hanging around her neck and you can’t feel her pull you in closer because she’s reaching farther behind your head of dark hair and tapping shoulders of random guys she’s never even met,

and when this happens I hope you run to the dingy bathroom and splash your face with dirty water and vomit up the words you never said because while you’re out drowning your heart in things I shouldn’t care about I’m here looking at the moon whispering how much I fucking love you,

and if you take her home I swear to God the moonlight will keep you awake no matter what time it is and you’ll watch it shine across your bedroom floor where we danced and laughed and I almost told you that you are my night sky,

and I hope the light catches your attention more than the sight of her would and I hope when you wake up all your remember is that roses are my favourite scented flower and you can’t escape the light of the moon.

I’m honestly over everything. I’m over this boy at work. I’m over him. I’m over how shitty life is right now. I’m over how upset I am because of how shitty life is right now. 

The PI I work with looked me in the eyeballs today and said, "if you get good results, we’ll publish it with your name on it. This means your westerns have to be perfect. Everything has to be perfect so it is publishable. This should be a good motivation for you. Also, I am going to stop helping you so much, I want you to struggle, so you learn on your own. You know enough that you don’t need me to hold your hand any more."

So you know what. Screw everything that is wrong right now. Screw beautiful men that are taken. Screw first loves that decide to ignore you and treat you like shit. Screw family problems that make you want to cry and pull your hair out. I’m pushing it all aside. I’m going to deal with everything else once I get into medical school. Right now, I’m going to focus on getting a paper published with my name on it before first year university ends. I want to for once feel proud of something that I’ve done on my own, all by myself.

I’m going to suck it up, ditch my emotions and do this.

Next experiments:

1. Finish last western. This one has to be beyond perfect.
2. siRNA knockdown of CLDN-2, plus TNF-α stimulus to see effect on proliferation.
3. Immunofluorescence and immunoblotting to confirm CLDN-2 inside nucleus of LLC PK1 cells.
4. Express CLDN-2 with nucleus localization signal to see whether increased localization of CLDN-2 inside the nucleus increases proliferation. Western, anti-PCNA to detect for proliferation.
5. Increase/decrease phosphorylation of CLDN-2 to see whether that increases localization of CLDN-2 in the nucleus.
6. Find out whether TNF-α causes a phosphorylation or de-phosphorylation of CLDN-2.
7. Explore relationship between TNF-α and ZONAB/ZO-1. 

he has a girlfriend.

life sucks.

i’m done.

so i talked to him today.

he goes to western.

first year medical school.

omfg. fuck me. literally.