GST, GST and more GST

Started even earlier today, i got to uni at 8:20am!!! Didn't feel tired which was a surprise. Derek was in a better mood today and more chirpy. He got his increased pay for his new job so he is happy. Got in and started with my GST binding method AGAIN! I'm beginning to feel really sick of this. Everyday its, bacterial lysate, assaying, GST binding, make gels, do SDS PAGE and sonification. I'm going mad!!!! Morning was uneventful. I even had time to have some breakfast which was nice. Although i have to say, bacon sandwich in SAF taste like cardboard. I'm not having that again! Bumped into Chui as well although she was in a rush. Then strolled back upstairs again. Wingkit was just starting his transformation or something. Then Derek mentioned to me that they bought some cake for Wingkit for his birthday. His is finally 21!! Old like the rest of us! Although he was reluctant about it. In fact he was moaning about it everytime somebody wished him a happy birthday! Talk about having a strop, Wingkit had one today, saying that everything that went wrong is all because of my fault!! Ah well, its his birthday, so i didn't really tease him.....well not a lot........fine, a little bit....... Well i only made him guilty once today!!! Muwahahahah!
Anyway, lunch was nice and the cake was better. We didn't quite sing Wingkit happy birthday, cos it would be too embarrasing since we were in the atrium. Mike treatened to holst Winks on their shoulders and run around the atrium. Haha, got Winks worried for a couple hours about it but they didn't do it in the end. poooo!
Anyway, it was back to my stupid bacterial lysate. In the end Derek suggested that i use the all of my lysate................. My gosh!!! Instead of messing with 200 microlitres of stuff, i was juggling 10 ml of the stuff. I should be getting quite skilled with all this....... actually i'm still hopeless. After a long time messing with the method, i finally managed to purify my enzymes (at least i hope i did) and was ready to load onto my gel. Although i cooked my sample a bit too long i was too absorbed with my acrylamide gel. Yes, looking for the tiny transparent wells of my small acrylamide gel is soooooo interesting that i was sooooooo enjoying myself....*heavy sarcasm...
Eugh, i have a lot of complaints today, i don't have a proper protocal, the journal about dichloro concentration of 1mM is absolute bullshit since it could only dissolve in 0.6mM and even then i had to heat it, i could only use one long pipette tip to load my samples onto my gel since each tip cost about 50p, i had to mix all the chemicals myself and some of the stuff i use are so lethal that i could get cancer if i expose a lot of it to myself, i keep having to autocalve stupid liquid media and conical flask, the 1ml gillson pipette always get mislaid, the glutathione has to be made fresh everytime i use it, i can't store my lysate in the freezer since i will ruin my enzyme and i wasn't told until yesterday, i have to leave BEFORE the supervisors, my finished gels are absolute rubbish since i didn't get any enzymes and most importantly I HAVE NO BLOODY RESULTS!!!!!!!! ........*heavy breathing........*weezing...........
Finished today EARLY for once and went computer room, chatted a bit and then went to the gym to jiggle my fat again at the treadmill. To top it all off, i found out that i gain 0.2 kilos....... ok, its not much but it sure didn't help with my self esteem..... and now i think i will go to bed........ *fuming with steam coming out of ears....!!

A long wednesday...

After a fitful sleep, woke up about 7:45am and forced myself to get up. After dragging myself to the bathroom to get ready, got dressed and went out. I didn't bother looking at the time, to me, i just thought, 'its early in the morning, i'm early'. Somehow in a daze i ended up in uni quite awake then and to my surprise, i was earlier than Wingkit and even Derek!! Woohooo! Anyway before i had the chance to pat myself on the back i started the day with a long winded GST binding method, to purify my proteins from my bacteria lysate. Needed to see if i managed to get some GST cos yesterday's assay was really really slow. The reading went up but slooooow!!! Anyway, back to faffing around with my experiment and messing with stupid chemicals and double checking my work, etc, etc. Somewhere in the middle of all that Wingkit came in and also realised that Rochelle and Paul wasn't in. They got the day off, lucky them!!
Felt really hungry by 11am but i was on the roll!! Unfortunately for me, both Derek and Mike were not and so i had to repeat my suggestions about my project to them several times! Man! That was annoying, all because they went out yesterday, celebrated and drank a lot. Oh, forgot to mention, Derek got a job offer, so thats why he went all celebratory. The GST bind took all morning and again about lunch time Wingkit announced that he will be finishing early again today. GRRRRR!!!! @#£$^&**!!!!! Why is it i never get anything time off!! I'm working myself to death here!!!!
It was about 1:45pm and i still havn't eaten. I was beginning to feel faint. I think its the dry air in the lab. Very bad for your health, especially when you are messing with lethal chemicals as well. I have no idea how much stuff i have been inhaling for the past 2-3 weeks. There is a definite increase of mutant cells in my lungs, if i do get ill, i'm going to blame all this on Derek!!!
Lunch lasted 10 minutes for me, then i came back up. Oh yeah, Steve also came in to check my methods. He practically corrected most of my stuff which was mainly rough anyway. He is sooo pedantic as well. Its scary, its like having a second Dr Lamb marking your work. Anyway, at least i know what he wants from me. A thoroughly checked work with all grammer and spelling mistakes searched for. Scary!!!!
I ran my SDS PAGE gel after lunch and managed to mess up half my samples. I had forgotten to heat my samples before loading and used 20microlitres of the protein ladder instead of 10microlitres. Hehe, it was expensive as well!! After a lot of swearing and stressed emotions, i got my gel to run and sat around waiting for 40 mintures. Oh, that was when i realised that Winks was still there. Instead of going home early, he had faffed around too much talking to people. Eugh, some people have too much time on their hands!!! After complaining how he can never find any journals relavent to his project, i disproved him in a matter of fact manner by finding some for him. It was not that hard actually....muwahahaha!!! Then i thought i would e-mail it to him. Little did i know that whilst i was opening my emails, Winks was so nosy that he stared at my emails. Of course whats in there, an email from Xiao with the title saying 'Wingkit's birthday list'. Ok, i could hit myself for forgetting that ppl might email me but still Winks was the nosy person who was looking at my mail when a normal POLITE person wouldn't!! ;P
According to winks however, he said he knew that we were going to do something but didn't know what and when. Also that he got suspicious when Xiyu mentioned that she in going to stay in South Kensington until Monday! So all he knows now is that something is being done but what and when is still unknown. After an ernest prodding at me about whats happening, he still couldn't get anything out of me, haha! It has wound him up as well, hehe!!
Anyway, back to the boring project, i started to make the phosphate buffer with the 1,2 dichloro 4-nitrobenzene...phew long chemical, it still didn't dissolve. So Mike suggested that we heated it. It did dissolve but then realised that i couldn't do the experiment since heat can speed up the original experiment and thus masking my results. Pooooooo! Daaaaang this stupid thing!!!!
Anyway, used the buffer anyway and did the assaying. Same type of results, even when i increased the bacterial lysate to 100microlitres. Ok, it was getting way to late, it was about 6pm now and Mike wanted to leave. So with no choice and feeling really tired, feet really sore and dejected, i cleared up and left. Eugh, was soo tired and now i'm just stressed. I HAVE NO RESULTS AND I WILL CRY VERY SOOOOOON!!!!

Hard at work...

Yep, was feeling a lot lazy for the past few days so didn't really bother to blog. Nothing much happened anyway, methods did not work again on Friday. The spec reading always gave a decrease, damn the stupid method. At least, i went out on Saturday to have dinner with friends. Then again on Sunday to Camden town to a party. It was my first time networking as well. Met a few people. However, my friend managed to attract two guys. I on the other hand stood like a vegetable to one side thinking, 'my feet is killing me, when can i sit down!!!" whilst my lovely friend prattled on and flirted. The guy didn't look to bad, although a bit on the thin side. Didn't like that. Second guy came up to us, or rather to my friend. Do i seem unfriendly or something?!?! i must chase people off or something. Hmmm....(on second thoughts no comments from u guys, Wingkit and Xiyu, i can tell what u would say!!!) anyway, he was an american and a lawyer to boot. Got offered a drink and accepted after some persuasion. Then this whole big conversation started. I have forgotten what we talked about, i think it was mostly about food. Anyway, when we finally realised the time, it was something like 10pm. Eeeek! My parents will kill me!!! So rushed off, but not before the guy asked us for our number. Lol, i can so tell that the number he really wanted was my friend's. Got home, faffed around and then went to bed late. Monday morning, i slept through 3 alarms. My gosh, it MUST have been the alcohol. Arrived at uni 30 minutes late. Then started the day with a meeting. Talked about our project. The discussion on my project went a bit long, i could tell Wingkit looked REALLY bored. But then mine was so much more complicated. It only lasted 30 minutes anyway. Then went back and made stuff to be autoclaved. Actually Monday was a nice relaxed day. Did some bacterial innoculation in the afternoon and faffed around chatting to people. Oh, before i forget, there was a small incident with a conical flask. I elbowed it by accident and it crashed to the floor. Thankfully, it didn't smash but it did chip at the flask opening. Hehe, i quickly hid the evidence in the bin and left the flask in with the others therfore seemingly making it look like it was already chipped before. Haha, fooled u people at the lab, muwahahaha!We all left BEFORE 5. Wow! It was the first time ever that i left BEFORE 5pm!!!! Went home and did nothing but watch chinese series. Then i went to bed at 9pm. Wow, havn't done that in years!!! Oh, apart from waking up at 11pm where my dad came into my room and asked if i went mad or something, cos i was sleeping so early. Hmmm, maybe thats true, going mad from our FYP!!!Tuesday....woke up at 7:30, thought....too early, sleep some more! 7:45am.....zzzzz.....8am...zzzzz...8:15...need..zzz..to..zzzz.. wake...up...zzzz....8:30am...oh crap...i'm going to be late!! Then rushed to uni. Horrible morning, so overcast. (this blog is getting loong) What did i do today...on yeah, did some calculations, more chemical mixing, washing, bacterial cell lysis, (i'm beginning to feel like a automatic robot!), more calculations.... Yeah, i bugged steve a lot for the calculations. I hope i wasn't being TOO annoying, hehe! Bothered Wingkit as well, to check my calculations. Actually today Wingkit bothered me first with his calculations. Although he was being very slow today, lack of sleep i think, lol. I had to explained it 3 times and he still didn't get it. When he finally understood, he went to check with Steve and lo, and behold, i was right 100%!! Whilst i was busy with my mounds of preparation work with my methods, Wingkit took a second to gloat at me that he will be finishing after lunch and therefore go home early. Pooo u!!!!!! I left 5:10, just when Steve had to leave. grrrrr!!!! (blog to long, too tired, speeding things up)I went to the gym, ran a bit with all my flab lolling around and then went home, showered, writing blog and now, i think i will sleep!!!

Method did not work!!!

Eugh, went to bed late and then woke up 5 minutes late, therefore i was 5 min late to uni. Sigh! It was sunny as well, but then i was so tired i didn't notice.
Got to uni and the first thing i knew was that i had to move upstairs to do my experiments but wingkit got to stay cos he got there before me! GRRRRR!!!! poo u wingkit. So, after wasting 30 min getting my stuff upstairs and to get used to the surroundings i found that it was not so bad after all. I get to have my own spectophotometer, there were three centrifuge machines, lots of pipettes and lots of SPACES!!!! hehe, the only thing it lacked was the company. Ah well, to make it up, i went downstairs to get some more stuff but also to make Wingkit really guilty. Hehe, and i succeeded, mwuhahahaha!!!
Awww, but then i felt bad so i stopped and went back upstairs again to face the stupid method. I did the assay i did yesterday and there was no reaction at all!!! It was 12 then and was ordered by Derek to go have lunch and do another method in the afternoon. So i trouped off with Wingkit to have lunch outside. Wow, it was really warm. I went to have ice-cream as well. Tried to coax Wingkit to have some too, cos i was feeling like a pig, but that did not work. I also noticed how thin he actually is when he keeps complaining how FAT he is. Ok, if he is "fat", then i must be OBESE!!!!
After chatting to Chin and Basma for a few minutes, we went back upstairs to face our project again. groan*
This time i did the GST binding buffer assay. Purifying the protein from the bacteria and then react it with the substrate (dichrlor stuff) and glutathione. I did the reaction and then found out that it did not work either. I also managed to bugger up the concentration/volume/measurement/counting/timing of the stupid method. Sigh, dunno if it really mess my results that bad or not. Then after Derek came up to check up on me, i realised something....i had forgotten to put the glutathione into the Reconstitution buffer!!!! CRAP CRAP CRAP CRAP CRAP CRAP CRAP*
I spent the last hour redoing most of the method which includes long minutes of contrifuging and shaking the mixture for ages. I got so sleepy that i put my head on the table and shook the tube in one hand. Just at that point i heard the door open and Dr Archer came through the door. I bolted upright and gave a sheepish smile. He laughed at me and asked if i fell asleep. I kinda looked embarrased and laughed in a silly way at him. My gosh why does this happen to me. The same thing happened before after lunch, but it was Dr Lamb instead. He also gave me a big smile and asked about my project. Told him it was going wrong and he replied that his second year results were good. I remarked how long that was, (actually it was only last year) and that i have gotten really old. He replied, "yes! You are 21!! I remember!" WHOA!!! That was really scary. So i laughed and said to him, yes i am getting really old. He said "yes you were all so young and innocent before." Ok, that was starting to get freaky....i kinda laughed and he left, but not before coming back and saying, "At least i'm much ahead in years, so you are fine in comparison!" .............speechless*..............
Sooooo today was another horrible day, i'm tired, restless, depressed and i banged my head on the table too! sniff* sniff*
Highlight of the day, i got a phone call from my lovely friend Naomi. After being comforted, i felt my day wasn't so bad afterall!!!

Bad enzyme day!

Today started ok. The weather was nice, not too cold or too warm. It wasn't raining either. The tube wasn't as cramped as yesterday so i managed to lean on one of these cushion thingy and stretch my legs out. Dosed for about 5 min and realised that there is a werid oriental guy in his suit sitting to my right staring at me. Hmm, maybe he thought i resemble somebody. Ignored him and briskly walked off at South Ken.
Got into uni and had a mini chat with Derek and Mike about my weird results from yesterday who then suggested that i should recalculate all the concentrations of my chemicals (crap*). Then i started the grueling work of calculating mols and volumes and concentrations. It went down hill from there. We found out that the concentration of 1,2, Chloro-4-nitrobenzene (1 mM as the paper suggested) was too high and that it couldn't dissolve in water. Hence the fluctuating results yesterday, since the substrated decided to crystallise out in the phosphate buffer. Then i had to adjust the concentration to 0.5mM. Eugh! I havn't oiled that part of my brian since A level chemistery. For some reason, i had a really bad mental block. My head couldn't get around the stupid mols and volumes and dilutions. That took most of the morning with me moaning at Wingkit and Derek about my incompetance. I think Wingkit felt sorry for me and came over to help me talk the concentration over, i probably looked really pathetic, haha. Wow! He actually remembered that i work better if i discussed things with people than just staring at a piece of paper!
After working out the concentration i was eager to start my assay. So added my GSH to my 1,2, Chloro-4-nitrobenzene, phosphate buffer then did 5, 10, 20 microlitres of bacteria lysate. After all that work and effort. I found that the results did not increase or decrease this time. But stayed the same within the region of +/- 0.001. i did triplicated of the three concentration of bacteria before i realised that it was 1pm. During that time, i also noticed that my sample of bateria lysate had a lot of things floating around (crap*), the unwanted cells had resuspended again. It hit me that there could be a possibility that the debri was masking my reaction. (crap crap*) So before lunch i centrifuged the bacteria and decided that i'm eating now.
The day was too nice, so grabbed a quick lunch from the MDH and sat on the steps of the queen's tower to eat. However, Marvin complained about the flies that were floating around and so we migrated into SAF again. After a werid conversation with Marvin, to which the contents can be guessed if those who know Marvin well can very well guess, I decided to brave the assay once more.
Alas, with no avail, the results were still bad. Instead I found that there was no increase or decrease from zero!!!! What happened to my enzymes and lysate?!?! (crap crap crap*)
After all that faffing around, Derek and I decided that we should regrow the bacteria again and lysed them for a second time. This was based on the fact that i grew my bacteria over the weekend and it may have produced a lot of proteases what may have broken the GST enzyme down again. (crap crap crap crap*)
Ok, seriouse tiredness now and low in spirit. Made my bacteria culture to incubate and then tidied up my workspace. Afterwards i left for the computer room to which as you can see i'm writing my blog. So tired. Today was not successful. Must rest well tonight. A looong day tomorrow....sigh......zzzzzzzzzzzz

Messed up results

So frustrated!!! Stupid method gave me stupid results that i can't use. What gives totally different results to triplicates of the same experiment!! RHAAAAAAA!!!
Ok, rant over! Well, kinda established that the problem probably originated from the fact the the pooey 1, 2 dichloro-4-nitrobenzene substrate won't bloody dissovle in water. Not at 1mM which is what i need. Crap!!!!! I'm still faffing with the method and i don't have much time left!!!!!
Today was tiring and yielded weird results. Eugh, too depressed! Going to watch something to cheer me up.

FYP first Blog

Finally, a new blog for my FYP. It was a bit late because i was being lazy and didn't feel like posting anything. But then decided that i'm missing out on stuff!
Now on third day of my FYP and i am as confused as ever with my project. :s Thank goodness all of my supervisors are really nice people. Although we are allocated different supervisors we can still ask the other. Although that does involve 5 min of confusion in the beginning before we managed to get on the same wavelength. Hehe, i think i do pester them a lot, they are probably really fed up with me.....having said that, i really don't understand what they are telling me. They did say to bother them when we need too. Haha!
Me and Winks postgrad is Derek Wallace. Techinically he is a doctor now but its confusing so we will say he is Derek. He does seem to have a lot of days off however. We had 2 days extra holiday before our project started. On friday he needed to go somewhere and left me and wingkit with Mike. Today, he went to the gym, then had a tutorial and then left early to buy a suit for his interview tomorrow. Which also means, me and Winks are left all by ourselves tomorrow. Ah well, thank goodness for Steve and Mike.
Experiment was slow however moving along steadily. Last week, i was making buffer only. My gosh, that was boring. However, the spec reading today was a bit weird. Instead of increasing value of absorbance value, i had a decrease. After stressing about it for 1 hour, then sitting around doing nothing, waiting for Wingkit for the next hour, i suddenly realised, i had forgotten to put one of my substances in! Crap, dang, sh*$, poo it!!!!!!!!!!!! Sigh! Ah well, at least i didn't break anything today except for one hairy moment when Rochelle decided to spill pure 8M (i think) down her front and called me in a panic to get Steve Cook, who conveniently was not present at his desk. After the initial shock, she calmed down, and took off her lab coat. A puzzle for you, how can the acid that is spilt on the lab coat not burn a hole in the labcoat but on her top instead (worn under the labcoat)?!?! Well, that really put me off working with such a strong acid.
Wingkit was sooo slow today. After the initial luck with his PCR, it seems to have run out on him. He kept complaining to me have he could have got a free day tomorrow, blah, blah, blah... Sorry Wingkit, hehe, no sympathy for you. I have done more work during the day running around checking my chemicals and machines with no opportunity of a day off at all!!!!! ;P
Oh crap! ICMUN now and i'm late!!! Well, i'm going to stop waffling and go.