Worst day in lab

Wednesday evening, we showed our poster to the lab members for constructive criticism and some proof reading. I had the least data and prof suggested 2 new experiments for me to do which I immediately did the transfection for that night. Today I woke up especially early so I could get a ride from my mum to school. I reached lab at 8 and started my cyclohexamide treatment, it was my first time doing it and I had 3 sets of 6 samples each. Then I cleaned and set up 4 gel plates during the 1 hour break to cast the gels I wanted to use later. Also added NaOH to the EGTA solution which wouldn't dissolve since yesterday night. One of the gels leaked and in between my cyclohexamide treatment I made another which still leaked. I harvested the 18 samples without much trouble, though it took quite long. Jsenior informed me that there was probably not enough cells after her experiments so I won't get to do a forward transfection to repeat my experiments. It wasn't anyone's fault but I was quite upset :( Close to 11am, I started harvesting my other sets of cells to do a nuclear and cytoplasm extraction, also a first for me. Finding the right person for the protocol yesterday took me so long and I was silly enough to insist on making my own buffer -- that got me home at close to 9pm. Today shall be better, I thought. I harvested the cells and followed the protocol until it was the 15 minute incubation on ice. I started to prepare 10% NP40 from the 100% and it just wouldn't mix properly with water. It clogged up my pipette and formed lots of bubbles when I tried to eject that disgusting blob of NP40. This repeated twice and I was on my third try when the NP40 just stuck to the outside of my tip and refused to budge. I was impatient and panicking because 15 minutes was up and I didn't want my cells to swell too much. I was asking FYP mate A for help, we discussed abit then I spotted FYP mate B and asked her as well. Then a FYP student from the other lab came over to kaypoh as she was walking by and started asking FYP mate A trivial stuff while I was in between them trying to figure things out.

Me aka overly attached fyp mate : Stop! I was asking her for help, don't talk now!

Her sounding a little offended: why don't you ask Jsenior?

Since J senior happened to be just around the area I asked for her advice too. Everyone was just telling me the same thing regarding the dissolving blablabla. FYP mate B and Jsenior were there counting the concentration and stuff. She asked to see the protocol which I didn't have printed out because my laptop can never connect to the printer. FYP mate A offered to print it for me and I could barely speak to tell her that the protocol was in my email, not my thumb drive because my eyes were full of tears and I was about to cry. I couldn't hold it in and started sobbing a little.

I didn't want to start new experiments this late in the FYP, within a hour of knowing the experiment, not as properly guided as I would have liked and well I was just hungry and stressed out and frustrated at the NP40. I knew I was going to miss lunch for the third day in a row and staying ridiculously late and I was so damn hungry having not eaten anything since 7am and being on my feet the whole time.

I took off my lab coat and went to get my matric card to go to the toilet but I ended up just sitting at my table to cry. Wah damn emo. I recovered, though not completely because tears started flowing again after they went for lunch and I was there washing my nuclear pellet. I had missed lunch so often and this honestly wasn't what I expected at this point of the project. It wasn't that I didn't work hard in lab, I worked so hard but got no data regardless or missed lunches or not. It just wasn't worth mistreating my body for. This whole fyp just isn't worth it. I don't want to do research. 

Before they went for lunch my sweet fyp mates managed to dissolve the NP40 and printed out my protocol while I stayed at my table blowing my nose. They also kept checking back on me throughout the day :') 

Anyways I went 'screw this gels, I'm going to eat' and only prepared my 26 samples for loading after lunch. I ran 4 gels together but to my dismay the 2 extended gels didn't run due to the faulty equipment and my dyes and protein had been sitting there for so long. By the time it was done with running and transfer, the ladder was almost gone and the dye and maybe proteins had leaked across the lanes. I cut the membranes and labeled them before putting all for a super short block of 30 minutes which would end at 8.15pm. 

It was only when I put the membrane strips into the primary antibodies that I realised a few of my blots were missing, I was panicking again and spent a good 10 minutes looking in the different dustbins. I found them later in the transfer tank which meant I didn't even take them out after the transfer was done. I didn't bother blocking and just cut and dumped them into the antibody tubes. 

I left lab at 9 and finished dinner at 10pm, even later than yesterday.

This post isn't meant to fish for sympathies, this is hands down the toughest period of my fyp journey and i want to keep a detailed record. I hope to look back at the entry in the future and think that I've grown stronger since this day. 

Quick update

I'm just studying for the 40% oncogenes CA tomorrow, leaving my poster and thesis aside. I managed to get more data, but need to repeat both experiments. I have so little data it's not funny... 

I also obtained my second shalnark card on hunter x hunter battle collections. In the process of getting a second card to enhance it. The picture on the card changes after enhancement so I'm all asdfgdowdksjsha

Here he is right now!

The state of my thesis

Happened to read what I wrote a few days ago and it felt like a joke. I'm so tired, I want to stop my experiments but I cannot because I had no results until the past week. Running so many experiments, desperately coming up with a poster to present to the lab ppl next week, writing the damned thesis so slowly and studying for a 40% CA next monday. I just want to stop experiments and focus on writing and studying. Or maybe I want to not write the thesis and presentation stuff and just do experiments all day long. I dont know man. The only motivation is that if all goes well, I'll have 2 more sets of data by this saturday. If not well I'll just keep working until time's up.

So tired

Ramblings

Living life to the fullest by going to lab every single day. At least I'm getting a little data to give me motivation for this last stretch. I have none for writing of the thesis though urghhhhhhh hopefully the urge to write will come really really soon :X

I can see the end of university... This is going to be the last few months of my life as a student and I'm feeling bittersweet about it all. I've been surveying if my working friends prefer studying or working......... It's a mixed response.. On one hand you can stop thinking about work after knocking off vs needing to do homework and mug after school, but you can't just take a day off whenever you like while you can skip lectures. 

But ya la.. it's part and parcel of life and there's no use thinking about which is better because there is no way to avoid graduation and working. 

Friends, I may have stopped posting about Hunter X Hunter but I'm still faithfully watching the new episode every week and sobbing and screaming at the development. I'm also following Hozuki no Reitetsu, Space Dandy, Yowamushi no pedal, Nagi no Asakura and Uchuu Kyodai, though there's a little back log because I was really busy last week.

Prop making has stopped and my mum's just screaming at me to clear the bags of the foam sponge thingy on my piano. But mum I can't, because I need those for making the prop and I have no time to finish it right now, so the materials must stay.

Met up with the YYYYs on tuesday and I spent like... all of my money LOLOL I was really down to dollars the next day because tuition stopped for a month and I will only be receiving the pay next week. Still it was a very nice meeting amidst chionging around in lab.

 Love you guys so much :)))

I'll just post some photos of my new favourite game after Kawai Pet Megu closed down-- LinePlay. I adore my character so much, this pretty little thing :))

                    

Isn't she sweet? She is capable of other expressions by the way, I just happened to screenshot her with the default expression. The second picture shows part of my lovely house which I painstakingly decorated. It's so pretty hahaha I'm like praising myself but I'm really proud of my apartment :D It's full of flowers, rainbow, more flowers and soft pastel fluffy things. I'm so girly hahaha it's hard to tell when you see me in real life hor? XD

                  

Oops I caught my cutie bathing hahaha and check out my garden inspired doorway :D

A closer look so you all can appreciate the loveliness ^__^

Updates~

I've fallen ill :( snacked on too many stroopwaffles from Mel and I woke up with a scratchy throat the next day which became sneezing fits... And now I have a leaky nose and a bit of a sore throat :/ 

Had a really busy week and I was out for dinner every single day, by choice of course but it was still so tiring.. Maybe that's why I fell sick :( I've been going to lab every single day including weekends.. Need a lot of preparations each time before I can start on the actual data generating experiments.. So time consuming haish

I watched hunter x hunter: the last mission movie on Wednesday!!! Managed to get the poster after asking the lady nicely if I could have one after the movie. The deal was 2 movie ticket to 1 poster.. Dunno why they were so kiam siap but my group of 5 ppl received 2 posters lol! I had to 'fight it out' among 2 other friends and I couldn't stand doing it the scissors paper stone way so I just went to ask for another XD fortunately the lady gave me a poster so by default that belongs to me ohohoho ^__^

          

I enjoyed the movie tremendously despite the questionable quality at the start and lame defeat of the bosses lolol so much feels whenever the ending song played in the movie ahhhhh.. It kinda reflects the current plot in the anime, with gon being so rash and worrying killua :X

Bought new shoes! 2 pairs for $36 and I love both pairs very much!! 

                          

So so happy~ :))) threw out my old pair wore the black pair today, it hurt my feet but all will be fine with some plasters! 

I also collected my Yang wig today! The previous wig I got was very pretty but too dense for the character and not curly enough. This one is just right, though I would like it to be thicker and less tangled haha oh wells. The ahoge is so cute!!!! Wore it together with the top which I got from auntie yesterday~

                                                

^.^ now I just need to make the skirt and finish up her arm props. And as usual try my best to lose the tan -__-

I might be happier without having to think about these

I've published this entry, reverted it to draft, tried to blog about something lighthearted, and now I'm posting part of the original entry again.

Holes on my arms

Every time I go for a blood test and the blood draw is unsuccessful, I always feel like I've lost a year off my life.. T________T

Doctor managed to find a nice fat vein on my right arm though he commented that it might slide around, making it hard for the syringe to enter. That was exactly what happened omg. I'm not faint when I see blood, though needles started to scare me a little after I pricked myself with one during one of my experiments last year.... I've had a very very bad experience drawing blood a few years ago and since then I've learnt a few more tricks to ensure a higher rate of drawing blood--- drink lots of water before going for it, point your fist down and rapidly clench and unclench the fist to make the veins pop.

Still, I felt my heart sink lower and lower as the doctor poked and poked around under the skin, he did it for a full minute and I was clenching my fist really tightly, and holding my breath as well cos he said to not move LOL. Didn't need to be so tensed, I was just scared I think. My muscles were starting to ache after the prolonged period of fist clenching and even after I got his approval to relax my arm, I could feel my head spinning X_______X I just knew it, that I was going to experience the loss of vision and ringing ears again -cries-

I politely gave indications that he should stop by going 'hmmmm' and 'aiyo..' so he took out the needle and loosened the arm band thingy. Slowly but surely my ears started ringing and vision was hazy. I can't quite remember what exactly happened but I was just sitting there with my head hanging down and trying to shake off the discomfort while he went to fetch another needle. He even patted me on the head LOL!! It didn't strike me as strange then but on hindsight I thought it was a very quirky but affectionate thing to do. Still,  sir I'm 23 years old and graduating in June this year please XD

He knew that I was feeling really bad and kept saying 'sorry, so sorry' as he felt for the vein and inserted the needle. At that point I didn't care anymore OTL I was feeling nauseous and looking for a dustbin in case I couldn't hold back my puke. Fortunately the blood started to flow hahaha I didn't even bother looking and just said 'that's great!' with as much enthusiasm as I could muster but I think it was around the level of a flopping half dead fish.

Again, can't remember what happened but I was really breaking out in cold sweat and so clammy that the plasters refused to stick to my skin. I just took them off but doc gave me a few to keep in my bag. He also asked multiple times if I wanted to lie down.. Tempting but I said no because if I moved from where I was sitting I might vomit lolol I must have sounded really pathetic when I said 'If you don't mind I'll just sit here for a while' at least twice because I was afraid  that I was disturbing him :(

Anyways the nausea passed after a while and I left the clinic and went back to lab.. sigh. Well it was memorable... the doctor patted me on the head :3