音時雨 ～Regentropfen～

I’ve mentioned before, I’m addicted to KOKIA’s newest release: “single mother／クリスマスの響き (Christmas no hibiki).” I especially love the first song, “single mother.”
I love the clean arrangement: it’s mainly piano, together with a soft drum beat and a silent guitar. They made a peaceful, warm, sweet, sad song, full of love, and so touching. It drove me in tears, once and once again.

Finally, I decided to write down and translate its lyrics, by myself, with my heart. (I also want to do other songs, but I’m busy now ^^
I can’t say it’s the best version; it’s just our rendering of this song. We’re not translators, anyways. XD

Omg, I haven’t updated for weeks! Busy days. XD
Just finished a paper assignment that’s due on Dec. 14, but I haven’t even started to work on another one that’s due on Dec. 8. Anyhow, the main reason of why I did like that is there will be a presentation that is needed to be based on the Dec. 14 one… OTL… >_>

So, why suddenly I decided to update here?
Hmm… some reasons. But the main one should be that I need to backup the way of output a file directory in a txt file I just know.

For me, I found my output file was a 1049kb txt file for the whole music folder (yet I still haven’t arranged my newly got music in another folder, orz… >_>)

Talk about others. Recently I finally tried OPERA UNITE, and it’s really nice and fun. I love it so much, especially the fridge and the whiteboard!!
Also, I decided to use FLAC format for lossless music since now. That means, I need to convert piles of my music… Fortunately, I can batch-convert in JetAudio~
KOKIA’s new iTunes release “single mother/クリスマスの響き” is so nice! Driving me in tears… T__T I WANNA COVER “single mother”!!!
I want to change my youtube channel, and link to my blog, under the tag “cover.” Because I want to get rid off all my previous (crappy) covers and RESTART! I also don’t think youtube has a good look like my blog, and I don’t like manage too many different places/comments… I like everything being integrated into one. Like google, although I’m not a chrome support. (hey, youtube belongs to google! lol)
I’m addicted to Bizet’s “Carmen,” and Mozart’s “Le Nozze di Figaro” again.

Okies… what else?
I need to know what happened in my lab work, it doesn’t work like what I expected…
I’m attending to a pre-Xmas party in the next week, so I have to write more, faster!!! Or I’ll miss the deadline! But I AM striking to write this blog post! LOL
Busy, busy, busy life. Haha~

At last… it’s already a page (in MS WORD), is it still a petit update?

At this noon, as usual, we had a lab meeting.
Steve didn’t show up until 15 minutes after the supposed-beginning-time.
“Where’s Steve?”
“Dunno. Eating?”
It was 14:00 already.

That would be possible. Steve, our supervisor, comes to lab at noon and work till night. Actually, at this noon, 12:00 around, a person came to our lab to look for him.
“He hasn’t come yet. Brushing his teeth.”
LOL!

“So… are we done?”
“Yeah, I think so.”
“Okay, let’s go back.”
LOL!

“I have an open hour for my TA at 3. I just sit there and do nothing.”
“You can bring your laptop to play games. And if there’s a projector, you can connect it to the screen.”
LOL!

We couldn’t start the meeting if he didn’t show up. Anyway, we talked by ourselves firstly.
A desk outside our lab is a bit large. Steve was considering on changing a smaller one. But there was no suitable ones in our department building…
“So, we can either wait for a new one, or just get rid of the current one right now. However, another way is, we may use a saw, get a piece off in the middle, and then meld the rest parts together. So it becomes smaller!”
“Hmm… sounds not bad. I think I can just cut the legs off, then re-stick them to the smaller-cut desk. You need not cut it off from the middle in this way.
“Hold on, isn’t the desk made of wood? Can you meld it?”
“No, it’s made of metal.”
“Oh, that will be fine.”

“But, what about to use glue but meld them together?”
“What kind of glue may be used in there?”
At that time, Steve came.
“You guys have started to talk?”
“Yeah, we were talking on the desk…” we told Steve about what we just said.
“Hmm… is the desk made of wood? So you can meld it?”
“No, it’s made of metal.”
“Ah, I see.”
LOOL!

Sometimes, the lab meeting can be rather funny!

I think I'm doing my project with a snail speed!
In the past month, since the last week in September, speak in a more exact way, I kept doing PCR day by day. I was glad to do it since I needed not to make more solutions, but I didn't mean I wanted to do the EXACTLY same thing everyday!
Gosh, my PCR didn't work for rather a long period!
Everything seemed alright. I didn't think I can even make mistakes on weighting stuff and put it into water according to the desired concentration. I adjusted my solutions to the proper pH. I did positive controls of my PCR using others' buffers. But, no matter what I did, my OWN buffer didn't work! What the hell!
I started to doubt everything, even the water I used.
Anyways,
Calm down, water wouldn't be a problem!
I read the recipe again. Hold on! It says “pH 8.4 for Taq buffer, if use pH 7.5 ones, both of Taq and PWO would work......”
Okay, that's the point.
I made some Tris-HCl in pH of 8.4. Then, PCR again...
Gosh, it worked perfectly...
The damn pH-issue cost me almost one month to solve it. Fine, I won't trust all protocols too much from now on... -__-
At least, I confirmed that I was not that bad on making solutions. XDD

Finally, time to try some new experiments. Transcription! Yay~
However, I was a bit nervous on it. I remembered the last time I extracting RNA. It was degraded totally. Fortunately, I only needed a short fragment, thus the degradation didn't matter. However, now it was all about RNA, would my RNA be safely extracted and kept?
I started my transcription. I added my PCR product and all other stuff needed. When I did PCR, all stuff was just DNA-grade. I knew it, yet I still used my PCR product directly in the transcription. Afterwards, I extracted the RNA. I loaded the final purified RNA onto a regular agarose gel. I put the gel into UV...
I saw bands!
My RNAs were all safe! Good!
At that time, I was a bit amazed by the “cleanness” of my lab. I used DNA-grade stuff in RNA-level experiments, and I got what I wanted! That's so wonderful!
Or, would I say it's only because RNAse hasn't found its way to me?

I've been in my new lab for one month. I did kinds of pre-works: making batches of solutions and buffers, and, if you wonder more, that was, washed piles of flasks, and them bake some of them at 200 centigrade for 6 hours; then filter the solution one by one and thus made them eligible for RNA-related experiments.
As a result, I filtered 15+ solutions, made about 20+ solutions, and 20+ buffers. My firstly-empty rack was full now.
Such work was not unfamiliar, but there were always kinds of new "single lab specific tips" waiting for me to know. No no aseptic rooms, no ultraviolet lights, no much of kits (in fact they only have one gel extracting kit), even no lap spoons, (the last one was for avoiding the cross-contaminant; actually I wonder if they will use alcohol burner when dealing with microbes... o__O ) They (soon the word "they" would become "we") make everything by themselves (ourselves).
"We buy nothing but only chemicals."
That was very true. Firstly, I thought at least there should have some essential buffers and reagents. But later I knew I was wrong. We have to make PCR buffers, dNTP mixture, the loading dye.
After done all of these liquids, I did my first PCR on September 28.
In my previous lab, we didn't use EB and used GreenView instead. Here we use EB (ethidium bromide) to dye DNA. Thus it could be the first time for me to contact EB. When I dyed the gel in EB-water, a senior guy here told me something important:
"Remember to change your gloves every 10 minutes if you're working with EB. EB will be able to go through your glove and then go into your body after contacting of 10 minutes."
Fine. I often change my gloves. No worries. I told him.
"Yeah, I know. Anyway..."
.
.
.
"I'm not joking!"
He said with a serious joking way of speaking and a funny emotion.
Wah! LOOL!
But I saw nothing but the marker DNA on my gel later.
My PCR failed? What happened? Because I did a positive control, we confirmed the problem was about the Taq polymerase soon.
"Hmm... I've ever doubted that batch of Taq. We need to re-purify some."
We need to re-purify some.
What? Not just adding the powder of Taq into its buffer? Re-purify?
I knew already that there's no ready-to-be-used Taq liquid, and I firstly thought they buy the power of Taq.
"No. Actually, in this lab, we start from zero. We make everything, that including Taq, Pwo, and T7. Here is such a lab where you need to do EVERYTHING."
I see, I see. That means, I can train myself very much, huh?
I'm not joking!
Hey, be more positive! Although it's a bit troublesome and time-consuming, I haven't seen the procedure of purifying some Taq polymerases and then use them in PCR. Nice chance!
Okies! I'm waiting for looking at the whole purifying of Taq, maybe Pwo as well! :3

I was always nervous about the TA workshop (from Sep. 2-3, 2009) after I knew it. First, I have no experience about working as a TA; second, I even had only a faint idea on what's a TA.
Anyway, no one can avoid time going by. That day came finally.
At the time of waiting for the beginning, I was a little bored. I could have put a book in my bag, but I didn't.
The ambience was nice. There were 8 round table in the room. We randomly sat around one and thus made up 8 groups. Some senior graduate students joined and helped us in the workshop. The one in my group, was a very nice girl. I liked her from the moment she introduced herself. Sat beside her, I felt less tensive.
In the first day, we talked about almost all basic thing of being a TA, and learned several cases. However, OMG, I still need to improve my listening skill! @__@
Yet the case study was very nice. Who knows what we would met in the future TA working!
In the next day, every new future TA would be giving a mini-presentation in the case of pre-lab. We had got emails from the workshop host about our own topics. I've ever seen so many TAs when I was taking my undergraduate lab courses. Some of them were nice. Although I know what they did in the pre-lab part exactly, but I had no idea on how to do the presentation. I chose to wait until the first TA workshop day. I thought I would get some hints from it.
Actually, I think I didn't get what I expected. I've got to do it by myself without any new hints from the workshop.
I organized what I was going to talk, and searched online for some data. Then I figured out some essential sentences. Okay, I had only 5 minutes to do the presentation, I needed no more nonsense but only the essentials.
So, I needed to do a practice.
Didn't I?
We not only needed to do the presentation, but also needed to re-watch our performance afterwards! Our prepositions were all videotaped!!!
Maybe that was what I most worried about. I'm always nervous in front of a camera.
Some people used overhead slideshow. I didn't. I really dislike the overhead project. Sometimes the text was just too faint to read.
Thus I didn't go performing my presentation until the break. I wrote my points onto the blackboard instead of on the overheads before I started.
I was nervous, and who wasn't?
Maybe I got too many pauses, maybe I failed to control my speaking speed, maybe my volume was a bit low, maybe I should have done some overheads... my presentation wasn't that bad after all! The reports from our instructors told me that.
Fortunately, I'll not be TA-ing in fall term. I still have time to improve myself. There're still lots of things to be learned. I still have no experience on so many issues of being a TA: communicating with students, marking lab reports, teaching others in an effective way, etc... I don't know what it will be at my first TA-ing course yet.
No one knows.
However, a good beginning is always great and full of hope.
Go on like it.
That's what I wish right now.

I landed in Vancouver on August 23, 2009, at noon 12:00.
It was a bit hot. The sunlight was soft. Clouds floating in the sky, light blue sky. My sight was covered by golden dust.
There was breeze, made it a bit cooler.
The Vancouver airport was like a maze. It was so difficult to manage to find the correct way I should go. Actually, for me that airport was werid. I'd rather like to say that I was in a hotel than an airport.
And fewer people were in the airport.
Even at the moment when I went in to my plane to Calgary, I still has no any real sense of that I was in Canada.
I sat down, and started to recall the whole time during getting my study permit.
That was rather a fun experience.

I think I've been so accustomed to queue and wait. Yet that queue was too long. All I worried about was whether I would be able go catch my next plane. It was only less than 2 hours left, but there were too many people to let myself believe I could be done in time in this queue. Anyhow, no time for me to wait like this. I explained my difficult to people in front of me. Fortunately, I got to move myself to the very front.
I went to the window. Before I said something, the officer said first.
“Hello!”
Hello. I said.
“How are you?”
My brain got in blank for a short while. What should I say? Then from the deepest part of my mind, the fading away memory of English I learned years ago revived.
“Fine, thank you.”
“Excellent.” He smiled.
Okay. Not a bad beginning.
Now, I have no idea on how long it kept. He asked me kinds of questions, but I've forgotten most of them. What I knew was everything went well.
“Good luck!” He approved my study permit finally and returned me my passport.
I left the window. There was still time for me to catch my plane.
Although it was hard to find my way to my plane, I had less worries on it: I've got my boarding pass, my plane would at least wait for me for a while. Then, the officer showed in my mind. Oh, my god, he was rather handsome!
I can still remember the moment when he said “excellent.”
What a nice feeling it was! :3
I planned to take some photos in Vancouver, but I didn't have the time to do it.
At about 14:00, The plane took off on time. I didn't miss it~

The other thing I felt lacking the sense of reality was that here in Canada, people use both English and French in official situations. Sometimes I got confused. XDD

The rest of the trip was far shorter than the first part of it. I had a nap, and arrived at Calgary about one hour later.
The plane flied through the white misty clouds. I like to see the sky that's full of clouds, and I also love going through them! Perfect weather it had! ^_^/
My trip ends in Calgary. Nothing troublesome happened. I was not over-tired, I didn't get jet-lagged, I was still sane with my not-so-good speaking English. I was standing in the world, the other side of the world.

I left my home city at 16:00, on August 23, 2009, and arrived in Calgary at 16:15, on August 23, 2009. Yet actually, the trip took me about 13-4 hours in all.
Yay~, I took only 15 minutes from one side to the other side of the world!