音時雨 ～Regentropfen～

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?

The black kitty was drawn by chance!
Happy Halloween!!

@pixiv

Snowing! It's winter now!

@pixiv

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

originally, this picture was supposed to be a girl sitting in a train. but it become like what you see now with no reason! XD maybe because it's autumn now? this afternoon I saw the ground was filled with golden leaves. ^^

@pixiv