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N4! And qPCRs...
So i picked up my N4 cert at the Japanese cultural association yesterday... And of course with the voucher I signed up for N3 as well. And yes, I got back my 55 bucks for the N4 registration as well. All because the examiners phone rang. Thank you miss examiner, whoever you are... :P
Now all that's left is for me to go and buy my book and Kino... speaking of which.. damn! I should have followed my parents out earlier... they were going orchard road, and I could have just walked over to Kino myself while they did their shopping... darrrrrnnnnnnnnnnn why didn’t i think of that earlier??? Damnn damnn damnnnnnn! Arghs. Oh wells. I didn’t want to go out because I did a little bit of stretching this morning and the scab over my ankle erm... broke? And puss is coming out again. I'm on leave tmr for the doctor's appointment so... arghs.
Oh yes. A correction to my previous post. MMDA airs in april and not last week. Two weeks break... nachospaprika made a mistake and since I hadn't watched VSA then i didn’t know any better :P I watched it last night though, together with last week's AnShi. Somehow I mistakenly moved it to my ext HD even though I've not watched it yet. I even uploaded the mirror to mega but i didn’t watch that episode. Lol just what was I doing? Downloaded the full episode of J's nep league as well and i shall watch it in a while. Was trying to do up my half done VSA review but didn’t get very far.
Hmm work updates... my monster comp was taken away from me :( Sadded. But it can't be helped. My sup didn’t like it too since it was purchased with our group's budget, but we aren't doing any computation that needs that much processing power, but the guys from the other groups are. Previously one of them have borrowed my comp a few times to run his program already, but then more people wanted to use me comp. In the end they just found another comp for me to use. No more 30 sec shutdowns and instant excel file opens. Aw. I do understand it though. The stuff they are running... it took over 6 hours on that monster comp. On the mac the guy was using, it ran for 3 days non-stop, and was still not finished. Now if i was running that sort of programs, I'd want a workstation to run it on as well. I would hate having to wait 3 or 4 days for one calculation to finish, when there's a better tool available to run the experiment, and said tool was 'not being used'. That monster was definitely an overkill for my work needs. Of course I enjoyed how stuff loaded instantly and all, but I'd probably be able to make better use of the processing power if I had that monster at home. Like how I'd be playing a video, doing a blog entry, while having the previous entry being uploaded... my poor mac can barely handle that now... and if it’s a PC i can just open up maple and afk online - simply because my computer can take it. Right now my home computer... arghs its dying. It took over 10 minutes to start up word the other day, when my sis asked for help editing her project work. The monster certainly set me new goals in terms of dream computers though. A 6k workstation huh? I'd like to have that at home someday :P Someday... in the distant future... hahaha. By then workstations will be even more powerful that it'll make the monster I used look like a piece of cake huh? XD
But yes, the troublesome thing is that I have to reinstall all the apps i had. All those customization stuff. This time round I backed up all my downloads, so I don't have to re-dl each of them... BUT i forgot I should have exported some of my settings as well. Like the phrase files for phraseexpress... Now i'll have to key in all my phrases again. Or my hotkey settings for some other programs... gah. I'll try and back up all the stuff into dropbox the moment i finish setting things up this time round. The three time I'm changing comp in 3 months huh? The 'new' one i got isnt that bad. 4gb RAM i think, couldn’t find the ram thing under device manager, but the windows about system page says 4gb. 3.3giga hz processor, duad core... which isnt too bad really, but its so slow compared to the 8-core monster. Still specs wise it beats my old mac hands down. 2.4ghz duo core, no wonder its so slow ne? Esp with the 10 thousand apps I run in the foreground and background. That new comp gave me a headache though. I keyed in a wrong value when setting up outlook, and then outlook kept crashing every time i opened it. Had to dig in the user account settings to change it to make it work. Then i installed the phrase express add on for outlook, and after that my outlook couldn’t open again. Didn’t have time to restart my comp and see if it's fine after that... bleah. Hope it'll be fine. Worst still, Tuesday I'd be busy in the lab with DNA extractions, and extremely busy too if they really did add more reactors as Santosh said they would. Normally I do extractions on Mondays but I'm on leave tmr so... :P So i'll probably only get around to tinkering with my computer on Wednesday.
My sup... he was really nice about it. He asked me to speak to him before I leave and it make me really worried for a while. I was thinking 'just what did I do wrong?'. And he dragged me out of the office to talk too. Turns out that he just wanted to apologise about the computer, and didn’t want our boss who sits right beside me to hear about it. She was the one who asked that I change comps after all. He got me scared for a while though... :P
Work load wise... for now I'm really ok. Might be bad when I have to do DGGE next time, but now I'm quite free actually. I can do the preparations for DNA extraction much faster now... i can prepare the qPCR plates really fast too... took about an hour preparing everything from scratch, that's much faster than the 1.5hrs i took just one plate earlier. even nanodrop readings are taking me lesser time as well. Last Monday i finished the dna extractions AND all the nanodrop before what... 5:30pm? That's a far cry from the 6 or 6:30pm it took me previously, and those were without doing the nanodrops too. Plus Monday i had to add in two samples at the last minute as new stuff came in. Yup, I am really pleased with my performance last week haha. I like challenging myself like this, to do things faster, rearrange my work flow so that i can be more efficient. That's why i always check the time when doing my experiments, so that i'll know exactly how long i need to finish a particular job. That way I can plan my work better. Of cos it goes without saying that the quality of the results must not be affected. You're doing things fast but not haphazardly. I'm pleased to say that for the most part my qPCR results are within 0.5 cycles of each other... except for those where there were problems with the DNA extraction and the duplicates showed a huge difference. That I don't know why... I think its really the low efficiency from the kit though. The yield is really really low compared to another kit (but its too late in the project to change, just a mere 3 months left anyway). And the whole thing is automated so its really not my fault. (Sample prep is really, really simple. No way I can screw that up!)
Hmm I like running qPCR plates. I quite like the plate preparation process. Hmm here's how the plate looks like. Pulled from a random website that google showed me.
You run the thing in a machine that looks like this:
The machine is pretty big, and from my previous work place when i asked for a quote, its around 80k SGD, which is more than my whole year's salary.
Hmm here's another picture of the 96 well plate, that shows the pointy wells of the plate.
Well the plates that I work with, unlike the first picture, is completely white. Totally, absolutely white. The letter and numbers are there, but its just raised lettering of the white plastic, so it can be a bit difficult to see at times. So what you have to do is to take a marker and mark on the side of the plates. Like half way though the columns, and every two rows. That's the usual way, but it depends on your plate layout. If you have a particular layout you may want to mark your plate differently and accordingly. Its just to ensure that you don't load the wrong stuff into the wrong well - which can be quite a trouble considering that its just a sea of opaque whiteness. Its hard to even see if you've loaded stuff into a particular well or if its empty. So yup, you do need a certain degree of concentration while loading that stuff. Oh and the pipetting. You're dealing with really small volumes here. For the template I'm using just 2ul. That's 2 micro liters. Which is like... this small little drop that you can barely see if you're like 30cm away. Hmm... You know 1 ml? That's like... a few droplets of water? Or like 1/500 of a 500ml bottle of soda? One micro liter is 1/1000 of that. Imagine. So yup I'm putting that little wee bit of water (plus other stuff) into each well. If you're not careful half your sample ends up on the tip and doesn’t get into the well, and there goes your results. Oh and you do need decently steady hands too. You don't need to be a surgeon, but if you go around jabbing into the wrong well you're going to contaminate everything. I like leaving each little drop of water on the side of the wells, and spin it down later. Just so I can tell which well i've added the stuff to and which well i've not. Also makes it easier to load the stuff in IMO. But that's just my personal habit/pref.
Hmm... is this stuff interesting? Of are you bored out of your mind already? :P In anycase I'll be stopping here, got videos to watch! XD Till next time!
no subject
I found your post really interesting!
I´m the only one in my lab doing the qPCR but I use a different machine than you. It´s true that requires lot of concentration and takes a lot of time at the beginning, but now that I have more practice, I really like it. It calm me down, and I like focusing in something time by time because there is a lot of noise in my lab!!!
So lets keep working hard both of us!!:) (sorry for my english)
no subject
Yeah it sucks when I can't get my technical duplicates to go below 0.5 ct value diff. Like just today! I paid extra detail when pipetting the samples but still there were like three wells with diff above 1!!!! Arghs.
It's pretty quiet in the lab here. But I like to listen to music. That keeps me concentrate more somehow :) so I'm listening to music a lot these days hahha. Hope my earphones don't spoil so fast!