I feel bad about saying how tired and stressed I am when I see the number of hours some of my batch mates are spending in the lab doing their FYP. These days I’ve been reaching just after 9 and finishing after 6. The other day I stayed till 8 because I couldn’t finish my experiments for the day. It’s nothing compared to those who stay till late in the night, but still, it’s a fact that I’m tired and stressed. I think I’m trying to accomplish too much with the remaining time I have left.

I have to repeat my Western blot tomorrow since the first one wasn’t good, and I realised that I’m missing a non-transfected control. I can deduce whether my protein is being expressed even without the control, but it’s better to have one for comparison. I didn’t realise this at all until Lawrence asked me today, when he helped me out with my Western. I shall just see how the blot turns out on Thursday, and talk to my prof about it.

Speaking of Western blots, I had such a tough time with it today. I finished developing my film at 12+ today, only to be told that it wasn’t good, and I should repeat the experiment with a 15% gel. And so I rushed to cast another gel, so that I would be able to reach the step where I can incubate with my primary antibody overnight. I casted 4 different gels, and they all leaked! By that time it was around 2, and I was so frustrated. Lawrence came to help me, and I used the gel he casted to run my SDS-PAGE. The ladder I had used for my first round had run out, so I used the only other protein ladder that was available. And then… when my gel was running halfway, I realised that I couldn’t see any bands of my ladder! Turns out that the ladder wasn’t a pre-stained one, and so I had to abort my experiment halfway. By that time it was 3pm, and I hadn’t had lunch. Also, I had initially planned to use the PCR machine at 1, but since I had to repeat my Western, I couldn’t do it then. At 3pm, I was really frustrated. I had planned my experiments for the day, but because the Western didn’t turn out well, I had to scrap all those plans. That pushed my schedule back, and now it seems even more unlikely that I’ll be able to finish my experiments, as the thesis submission date is drawing near.

Excluding this week, there are 7 more weeks to go until the thesis submission date. In the ideal case where I get 4 weeks to write my thesis (and prepare my poster), it means that I only have 3 weeks left for experiments. In that amount of time, I have to repeat another round of transfections, which takes a week. I probably have to culture cells for a non-transfected control as well. And then it I need to do another Western that will take me 2 days, and that’s assuming that it goes well and I don’t have to repeat it. And if I have to… that’s 1 week? I still have 4 more genes I need to clone. I’ve PCRed 2 of them, and I’m trying to amplify 1 more in larger volumes. As for the last protein, I’ve run out of the cDNA template, which means that I have to make some by myself. The problem is that (1) I have absolutely no idea how to do it, and (2) I highly doubt that I have enough time, especially when my experiments seem to fail the first time I attempt them. Assuming that for the remaining 3 genes, I manage to get them cloned on first try, and taking into account the time needed for the sequencing results to come back and for me to do a midiprep… I might just be able to squeeze in a round of transfection and luciferase assay. I say MIGHT, since nothing ever happens according to the way that I plan it. It always takes longer than expected.

It’s been a pretty rough 1 week plus, with so many things not working well. I’m really hoping that tomorrow my experiments will work more smoothly… Gotta hang in there.

It’s almost 11.30pm, and I really should be getting to sleep soon so that I won’t feel tired tomorrow. But I can’t sleep… I don’t feel the slightest bit tired at all. Right now, I wish that tomorrow won’t come so soon. I don’t feel like going back to school yet.

The past week hasn’t been an easy one. I was very busy with my project, trying to do so many different things at a time. I felt flustered, stressed, tired and worried, wanting to let all those negative emotions out yet trying my best to keep it in. I try to smile and get through each day, but it was really tough.

This week doesn’t look as though it will be that much better. Still pretty busy, and I’m quite worried about some things as well. Mood’s still not too good, to be honest. I keep telling myself that things will probably be better after this week, when I get another small part of my results, and I can relax a little. I’ve been telling myself to be positive, because who wants to be around someone who’s unhappy? But sometimes I just need to let it out somewhere; I need a listening ear and some support as well as understanding from others. Having said that, I’m really thankful for the people who have encouraged and helped me along the way. Without them, I wonder how I would have gotten through the past week.

As I get nearer to graduation, the future seems to feel more and more uncertain. So many things are changing, and it feels scary. It’s like I’m trying desperately to cling on to what I have now, not wanting things to change, but knowing that they will. Trying to hold on to what’s familiar to me, but knowing that I can’t hold on to them forever…

I’m like a mixed bag of emotions these couple of days, but I’m sure that things will look better soon. After all, if things are at their lowest, the only way they can go from there is up, right? I shall try to remind myself of that.

I came across scans of this online, and so I decided to take a look at what sort of rooms were featured. It’s pink, pink and more pink, but I did see some rooms that I liked.

Went to The Fullerton Hotel for their Chocolate Buffet last night! Being Valentine’s Day, lots of people made reservations, and there were many extra tables set up for the night. Quite a lot of people didn’t turn up though, but that means less people around and a little more privacy.

These days, I’m usually too tired to do anything after I get home. It’s always the same cycle of waking up, going to lab, coming home, having dinner, relaxing for a while and then going to sleep. I get 8 hours of sleep almost everyday, but it doesn’t seem to help.

Project wise, I’ve been getting some results, but they’re not conclusive. I might need to do drug treatments to see if it can boost transcription activation, or express another transcription factor to see if the problem lies in the system itself (I’m trying to express a Plasmodium protein in a mammalian cell line, and there’s a chance that the mammalian transcription system might not be compatible with Plasmodium proteins.). My greatest concern now is whether I will be able to finish my project in time. Apparently we aren’t graded based on whether we get results or not, but who doesn’t wish to have results to show? And if we decide to go with trying to test if the system is compatible, I think it would be really good to have that result in my thesis. It would really give better weight to my current set of results.

The thing is, if I need to express some other proteins, I estimate that the PCR, cloning and all that would take me 2 weeks, and that’s assuming nothing goes wrong. And then I need 1 week to transfect my cells and do the assay, and I need to repeat the transfection to see whether the results are consistent. I have 9 weeks more to go till my thesis is due, and minus the time I need to write my thesis (I’m hoping to have 3 weeks at least), I might just have enough time to get another set of results. Might being the keyword here. I’m starting to feel the heat…

All I can do now is to take things one step at a time and see how it goes. Shall see what will happen next after I meet up with both my supervisors next week.