One more time, One more chance

秒速５センチメートル theme song by Yamazaki Masayoshi :

ONE MORE TIME, ONE MORE CHANCE

The video plain teases you with all the hints of what's going to happen in Chapter 2: Cosmonaut and Chapter 3: Byousoku 5 Centimeters. Makes me want to scream, "When is the DVD out???"

No better song to complement this romantic masterpiece apart from Yamazaki's heart-rending rendition of 'One More Time, One More Chance'. The title of the song speaks itself.

Here's the lyrics to the beautiful theme song in Kanji. Romanized version can be found on Gendou.

これ以上何を失えば　心は許させるの
どれ程の痛みならば　もういちど君に会える
One more time 季節よ　うつろわないで
One more time　ふざけあった時間よ

くいちがう時はいつも　僕が先に折れたね
わがままな性格が　なおさら愛しくさせた
One more chance　記憶に足を取られて
One more chance　次の場所を選べない

いつでも捜しているよ　どっかに君の姿を
向いのホーム　路地裏の窓
こんなとこにいるはずもないのに
願いがもしも叶うなら　今すぐ君のもとへ
できないことは　もう何もない
すべてかけて抱きしめてみせるよ

寂しさ紛らすだけなら　誰でもいいはずなのに
星が落ちそうな夜だから　自分をいつわれない
One more time　季節よ　うつろわないで
One more time　ふざけあった時間よ

いつでも捜しているよ　どっかに君の姿を
交差点でも　夢の中でも
こんなとこにいるはずもないのに
奇跡がもしも起こるなら　今すぐ君にみせたい
新しい朝　これからの僕
言えなかった「好き」という言葉も

夏の想い出がまわる
ふいに消えた鼓動

いつでも捜しているよ　どっかに君の姿を
明け方の街　桜木町で
こんなとこに来るはずもないのに
願いがもしも叶うなら　今すぐ君のもとへ
できないことは　もう何もない
すべてかけて抱きしめてみせるよ

いつでも捜しているよ　どっかに君の欠片を
旅先の店　新聞の隅
こんなとこにあるはずもないのに
奇跡がもしも起こるなら　今すぐ君に見せたい
新しい朝　これからの僕
言えなかった「好き」という言葉も

いつでも捜してしまう　どっかに君の笑顔を
急行待ちの　踏切あたり
こんなとこにいるはずもないのに
命が繰り返すならば　何度も君のもとへ
欲しいものなど　もう何もない
君のほかに大切なものなど

～みはる～

T_T

It's back to no-colony days.

TT_TT

～みはる～

秒速５センチメートル - 第１話 桜花抄

やっと来ました～！
新海誠の秒速５センチメートル - 第１話：桜花抄


The Japanese animation movie most anticipated this Spring - Shinkai Makoto's Byousoku 5 Senchimetoru. The first part of three, entitled Chapter 1 : Oukashou, delivered what the trailer had promised - top notch animation, excellent voice acting, gorgeous graphics, and beautiful music.

For some (maybe obvious) reason, animation movies are of higher quality than animation series. It probably has to do with the budget and direction, but animation movies have such fluid scene transition and fine-tuned animation it blows your mind. My first exposure to such superb quality animation was when I watched Miyazaki's Spirited Away. The animation of the characters and the scenes were very real. And I loved the graphics and colors. Although the plot was more supernatural, the voice acting was pretty good and dramatic.

Take Miyazaki's Spirited Away and multiply the quality by 5 times - you get Shinkai Makoto's Byousoku 5 Senchimetoru. While I do love Spirited Away, the first chapter of Byousoku 5 Senchimetoru really blew my mind. I love love love the way Shinkai-san animated the falling sakura petals and snowflakes... the sceneries throughout the Oukashou chapter were simply surreal. The animation is simply very REAL, especially in character-less scenes, i.e. the moving JR train, the falling snow at the train station, the gust of spring breeze carrying falling sakura petals, the reflection of lights against the classroom chairs,.... I could go on and on. I have never experienced such gorgeous graphics and animation. And this is only the first part of three.

I must also mention the plot and the direction of Oukashou.
In one word : brilliant.
The direction is very good because you can feel the frustration welling inside Takaki-kun when his train is delayed at every station as the snow falls mercilessly. You can feel his dismay as the snowstorm stood in his way and time passes by him in his journey to meet Akari-chan. You can feel the pain crushing Akari-chan when she relayed to Takaki-kun that she had to transfer to Tochigi. Most of all, the excellent direction makes the audience wonder if the deep bond and love Takaki and Akari had for each other could stand against the growing physical distance between them. Will they see each other again? Will time erase all special feelings they now held dear? Will time and distance drift apart two individuals with such strong affection for each other?

第３話　秒速５センチメートル holds the answer.




Please visit 秒速５センチメートル for more gorgeous images and trailers.

～みはる～

PhD... maybe?

"You have been short listed for an interview for our PhD course on 28 March 2007"

I mentally and emotionally went through a rollercoaster when I saw this email yesterday. I started off:

...feeling happy because I've made it to the interview stage, then
...feeling excited about the prospects of doing a doctorate, then
...feeling nervous thinking who the interviewer would be, then
...feeling unsure if I really have the means and time (and money!) to do PhD, then
...feeling restless deciding (already!) between PhD and a job, then
...very BLUR because I thought 28th March is NEXT WEEK.

Had my colleague, A, not remind me that 28th is tomorrow, I would have missed the interview altogether. And come to think of it, then I would not have so much to think about. Hm.

I think I think too much.

Wish me luck for tomorrow.

～みはる～

Plate of Happiness

My plate of happiness:



Yes... this is the plate of my happiness. I have a library!!!! *tears of joy*

I have been working almost 20 days in a row, in frustration I might add, just to get this baby. I have come to the conclusion that constructing a bacterial library is anything but simple. The procedure is deceptively straightforward, but I had my downs (and ups!!) in constructing this library since January. Considering the deadline to my thesis is one month's time and my project was not going anywhere without one bloody library up, I was already at my wits end doing tons of complicated optimization experiments. But one look at this plate... I'm drown in delirium!

For the past 2 weeks I was either scowling at the computer, scribbling calculations on recycled paper, or doing transformation. Every night my colleagues would see me at my bench pipetting like a mad girl and transforming bacteria, that I have come to proclaim myself the E.coli 'transformer' of the lab.

After 2 whole weeks mind-boggling optimization, I finally constructed the library for the 8th time with all the optimal conditions. When I checked the plates in the incubator the next day, my eyes could not believe what I was looking at. I bounded out of the incubator room to my colleagues in the lab and screamed, "I have my library!!!"

I must have looked really silly dancing around in the lab in delirium, while my colleagues laughed and congratulated me for obtaining the pretty pretty precious colonies. The joy of a successful experiment!! I have never felt so happy in my FYP-life.

However, I kept my feet to the ground, and thought that it would be best to check the reproducibility of my concept. Feeling incredibly motivated, I constructed another library using the same concept and cells. And guess what I saw the next day??

EVEN MORE COLONIES!!

I have a feeling I would be delirious for the entire weekend.

To give you an idea why I should be delirious, compare the before (library done two weeks ago) and after (library done two weeks later):



Before



After

I'm in cloud nine. 幸せだ～～！

～みはる～

桜日和

Bleach ending theme 10 - Sakura Biyori -

桜日和
Performed by Hoshimura Mai

十六で君と逢い
百年の恋をしたね
ひらひらと舞い落ちる
桜の花びらの下で

逢いたくて駆け抜けた
陽のあたる急な坂道や
公園の隅二人の影は
今も変わらぬまま

君と僕と“桜日和”
風に揺れて舞い戻る
まるで長い夢から覚めたように
見上げた先は桃色の空

好きでした好きでした
笑顔咲き染めた君が
僕だけが知っていた
右側やわらかな居場所

桜の下の約束
「来年もここに来よう」って
何度も確かめあったけど
今も果たせぬまま

君と僕と“桜日和”
風にそっと甦る
君も今どこかで見てるのかなぁ
あの日と同じ桃色の空

追いかけた日々の中に
刻まれた足跡は
何よりもかけがえのない宝物

君と僕と“桜日和”
風に揺れて舞い戻る
とめどない想いが溢れ出して
涙がこみ上げた

君と僕と“桜日和”
風に揺れて舞い戻る
まだ見ぬ未来を胸に抱いて
見上げた先は桃色の空

Lyrics from Gendou
Images from Random Curiosity

～みはる～

Allegro Cantabile

Nodame Cantabile (anime) opening theme

Allegro Cantabile
Performed by Suemitsu and The Suemith

88の場面の中に
散らばった音掻き集めた手
言葉にならない思い並べた
五線譜の上 追い越した日々

交差していく 三度を重ね
ここに響かせてー

歌う様に真っ直ぐに
歌の上を伸びていく
他には何も出来ない
時の全て預けながら
自由に解き放たれた
素直に駆け巡る指
終わり無き クレッシェンドは深く
誰かの元へ向かう

目の前に開かれた世界
繰り返す新しいプレリュード

88の夢物語、落とした休符、変わるハーモニー
静かにそっと整えた息…
在るべき形 取り戻すまで

掻き消される 三度を重ね ここに響かせてー

歌う度に信じていた 歌の側を外れていく
インテンポで躓いて
縺れていく即興の中へ
引き返す道が消える 動かされる波が近く
失った声のメロディ 誰かの元へ向かう

目の前を閉ざしていく扉 悲しみは
積み上げられたエチュード

意識もなく尖らせて 掴んだ肩振り切った
いつまでも忘れないで 歌う如く心のまま
歌う様に真っ直ぐに 歌の上を伸びていく
終わり無き クレッシェンドは深く
誰かの元へ向かう

ステージの 端のピアニストへー
生きる事 こんな歌の全て
今この目の前に開かれた世界
繰り返す新しいプレリュード

Lyrics from Gendou

～みはる～

When Brahms meet Paganini... and Kang

Every Wednesday of the school term, a series of Lunchtime Concert is held in one of the lecture theaters in my campus. The university, in collaboration with an external sponsor, introduced this Lunchtime Concert programme as a move in exploration of (musical) arts. Invitation emails are sent to the campus community every week, for performances such as piano recitals, string quartets, traditional instrument performances, and all sorts related to musical art.

This Lunchtime Concert series had been going on for a couple of years now, and until today, never had I experienced it once. Either I did not have the time, or I simply do not bother (usually the latter :P ).

However, thanks to the influence of Nodame Cantabile drama, I had come to enjoy classical music. If I had the chance, I would like to watch a Philharmonic Orchestra performing Beethoven's Symphony No.7, or Rachmaninov's Piano Concerto No.2. One of the piano performances in the Nodame Cantabile live-action drama that piqued my interest was Yuuto-kun's playing of Brahm's Variations on a Theme by Paganini. Although he was a meanie in the drama, the technique was very impressive. Watching the scene, I could only think, "Wow.." The piece appeared technically difficult, but the sound was beautiful.

Thus, when I saw the invitation for the upcoming Lunchtime Concert was a Piano Recital by Juyeon Kang who will be performing Brahm's Variations on a Theme by Paganini, I was determine not to miss this one and only piano recital. Die die, also must go.



The performing pianist, Juyeon Kang, is a widely-travelled and highly sought after concert artist. She has a string of impressive credentials, and has been described as a pianist of "formidable technique and exquisite sound". I admit to having know nothing of her, but I definitely did not want to miss a live performance of Brahm's Variation on a Theme by Paganini regardless who the performing pianist is.

However, my gel electrophoresis ate up 15 mins of my time, thus when I reached the lecture theatre, Juyeon Kang had already begun her performance of Hadyn's Sonata in C. Although it was only an hour long, the recital was fabulous. I thoroughly enjoyed all the pieces, especially Grieg's Wedding Day at Trouldhaugen and Brahm's Variation on a Theme by Paganini. The live performance of Brahms was beautiful. There was so much energy and spirit in the performance, that I could feel my pulse quickening at the crescendoes. Watching her play, I was impressed with the technique. Brahm's Variation of a Theme by Paganini is said to be one of the most virtuosic works of Brahms, and it is technically very difficult to play. Juyeon herself said that she took 3-4 years to master it! And she mastered it well, because I love her performance!

Now I'm just waiting for another pianist to hold a lunctime recital to perform Rachmaninov's Piano Concerto No.2 with a full-fledge orchestra......

PS: Did you know that Paganini is a historical violin virtuoso who was capable of playing three octaves across four strings in a hand span? That is considered an impossible feat even by today's standards and it was said that Paganini traded his soul to the Demon for this violinistic technique... In contrary, I think he's got some kind of genetic diseases that gave him long and flexible fingers to become such a virtouso.

～みはる～

合格できました！

去年した日本語能力試験が合格できました！

Yep, I passed my JLPT 3!!!
うれしい！！！
I have just received my score report telling me that I had passed the exam with a total score of 93%! This is quite unexpected! I was confident of passing, but never thought it would be with such a high score. I mean, afterall, I didn't have a proper preparatory course (one day of cramming, that was it!) and I did alot of guessing to the many questions which I have no clue at all. I must have been very lucky at guessing on the exam day, because I scored a 96.5% for my grammar. I totally sucked at grammar!!

Of course, I wouldn't have been able to even pass the exam without the excellent tutoring and dedication of my two Nihongo sensei. They have been the best teachers I ever had!
藤田先生、リー先生、本当にありがとうございました！

まあ、これから日本語の勉強もっとがんばります！
日本語が上手になるために、ちゃんと授業を出て、もっとたくさん事を勉強します。
もちろん、もっとたくさんアニメとドラマを見ることも大切なんですよね　：P
これからがんばります！！！

～みはる～

Bleach 117 - 初めの舞、月白

I've been waiting for this to be animated for a long long time:

舞え、袖の白雪！

image from Memento

～みはる～

Cooking Hazards

For the past few weeks, I have been preparing homecook food, and in the process, have been exposed to various cooking hazards. Let me share with you my top two hazards, and the solutions I have devised.

Top 2 hazards of food preparation:

1. Worm in vegetable

• Hazard:

When cleaning the rosette-bud lettuce called locally 'Eu Mak', beware of elastic earthworms hiding behind the ridges of the leaves. These eyeless, boneless, and limbless creatures are usually found burrowed in the juicy stems of the leaves and will wriggle fiercely when provoked.

• Solution:

Hold the rosette-bud vege an arm's length away (you don't want worms wriggling at your face), and use a knife to cut the stem of the bud. Let the leaf drop onto the sink (you don't want the worm to crawl onto your fingers). Check the leaf to ensure no worms are present, then turn the leaf over to check again (you don't want worms wriggling into your esophagus). If something maroon, long, elastic-looking, and is wriggling, pick the leaf with a forcep and dunk it in a yellow biohazard bag. Repeat the same for every leaf before rinsing with clean H2O. Place the yellow hazard bag in the incinerator.


2. Worm in vegetable

• Hazard:

When peeling sweet potato skin, beware of tiny, white worms burrowed in the tuber. Do not be fooled by the small size nor innocent white appearance of these wriggling things; they can be a menace to your sweet-potato-dessert preparation. These evil wrigglers can be found munching carbohydrates under the surface of the tuber skin, and will cause cardiac arrest to individuals peeling the tuber skin with a common kitchen peeler.

• Solution:

When washing the tuber under running tap water, check the surface for labyrinth-like marks on the tuber skin. If you see a lot of localized S-shape markings, immediately dunk the tuber into a yellow biohazard bag. Double-bag the biohazard bag, and throw it into the incinerator. Should you fail to notice these markings, and come face-to-face with something white and wriggling in the tuber, obey your reflexes, release the tuber from your grasp immediately (you don't want worms wriggling in your hands), throw boiling hot water over the tuber (you don't want worms crawling up the sink) and dispose the tuber in a yellow biohazard bag using a pair of thongs. Incinerate the biohazard bag IMMEDIATELY.



I think you get my point.

PS: Biohazard bags are optional and are replaceable with PP-type plastic bags.

～みはる～

SUEMITSU & THE SUEMITH - Allegro Cantabile

When the anime version of Nodame Cantabile premiered last January, I was quite apprehensive as to how JC Staff will be managing the music aspect of the animation. Nodame Cantabile is a classical music-themed piece of artwork, and the live-action drama had adhered to this theme fiercely. The opening and closing themes of the drama were Beethoven's Symphony No. 7 and Gershwin's Rhapsody In Blue respectively. The use of these two original classical music recordings complemented the classical music focus of the drama, and as a result the significance and beauty of classical music was very well delivered in the drama.

However, almost all anime opening and ending themes either fall in the J-pop or J-rock basket, thus I was afraid the use of such contemporary recordings would not only ruin the classical music theme of Nodame Cantabile, but would simply sound plain weird. I mean, imagine rock music and Beethoven's Piano Sonata Pathetique together... it's just plain weird.

However, when I saw the opening of the first episode of Nodame Cantabile, I was pleasantly surprised by JC Staff's ingenuity. The opening theme is an upbeat song written and performed by Suemitsu & The Suemith called, Allegro Cantabile. I thought the track was very creative in that it was sort of a reference to an important track in the story, Nodame's lively rendition of Beethoven's Pathetique - Adagio Cantabile. The tune is quite infectious and to top it off, the opening sequence is very attractive (although quite reminiscent of Honey & Clover II's Fugainaiya opening sequence). They had also cleverly incorporated snippets of Beethoven's Symphony No.7 in the opening theme.

The Allegro Cantabile single has since been released by Suemitsu & The Suemith on 21st February. I had not listened to Suemitsu & The Suemith before, so I took it from the name that it is either a band or a group of 2. To my surprise, Suemitsu & The Suemith comprise of only 1 guy, Suemitsu Atsushi !!!　O.O

Anyway, I find the song pretty good and the PV is quite cool too. Watching it, you can't help wondering what the hell happened to the grand piano. Try throwing Nippon black paint over an ivory white grand piano?

Here's Suemitsu & The Suemith for you!

By the way, do you think Suemitsu-san actually played the song with such slippery fingers? There's no way I could hit the right keys with black paint all over my digits.

～みはる～

あいぞめ

Jigoku Shoujo Futakomori Ending Theme - Aizome -

あいぞめ
Performed by Noto Mamiko

あさきゆめみじ　永久に　嘆きもせす

月の光　心　照らし出す
燃ゆる花の舞　道しるべ

絡みつく　あやまちの詩　胸をしめる

果てぬ川に手を挿し　流そう
想い　詰めた言の葉
藍に染めて

すれ違いが　心もろくする
居場所もないまま　風は吹く

赤い道　足取り重く　闇に向かう

一度　流せば　二度と帰らぬ
指が　まびく運命
藍に染めて

いくつ　明けない夜を　重ねて
やがて　藍の嘆きも
消え逝くのか

果てぬ川に手を挿し　流そう
色は匂へど　いつか
散りぬるもの

藍に染めて...

check out Memento for PV images

～みはる～

Anime: Kanon Ep 21

Since last autumn, I have been following Kyoto Animation's remake of the acclaimed anime 'Kanon'. The main plot of Kanon revolves around a high school guy named Aizawa Yuuichi and the snowy city of mystery and miracles in which he is currently residing in the present time frame. Yuuichi had lived in the city as a kid 7 years ago, but moved out with his family during his elementary years. Fast forward 7 years later, Yuuichi returned to the city, but mysteriously, he had little recollection about the city, nor any memory of his life in the city 7 years ago. However, since moving back to the city, he met a string of girls, who slowly trigger the return of his memories in the city. And it seems that we will soon find that his memories about the snowy city 7 years ago was a traumatic one.

I had not mentioned anything about watching Kanon, nor review it since its premiere on Japanese airwaves last autumn. Apart from not having the time, I find the story pacing very slow, and sometimes quite mundane. However, I still follow the series every week not only because the first production was a success (storywise, and also production wise), but particularly because I quite like Kyoto Animation's work. One of KyoAni's production which I liked best was the animation AIR. I love their artwork, fluid animation and soundtrack. The way they shoot the scenes create a subtle, yet incredible impact. Furthermore, this remake of Kanon boasts an impressive seiyuu (voice actor) cast: the male lead, Aizawa Yuuichi, is voiced by one of my favorites, Sugita Tomokazu. Other seiyuus include Horie Yui (Tohru of Fruits Basket, Naru of Love Hina), and Seki Tomokazu (Nobu of NANA, Chiaki of Nodame Cantabile).

Now, I admit that I had been watching the first 60% of the series impassionately. I do enjoy listening to Sugita Tomokazu's acting, especially the sarcastic jokes and dramatic demeanor. While each arc of the series is decently interesting, they were not over-the-top exciting. There's sort of like a degree of mildness and calm in the way KyoAni presented the story. However, watching the ending segment of Kanon episode 21 took my breath away. The way KyoAni introduce such a shocking turn of event in the calming pace they had adopted all this while is excellent.

Let me just highlight 2 particular segments I liked best in this episode:

1. Nayuki and yuki-usagi (snow bunny)

While I understood that Nayuki had all this while liked Yuuichi, it made me very sad when they showed the link between Nayuki's unhappy memories of the snow bunny with Yuuichi. I did not expect Yuuichi to have delivered such a harsh blow to her sincerity 7 years ago, and yet she can still like him after such a hurtful gesture. The scene showing Yuuichi's memory of the shattered snow bunny and the relevation that it was to Yuuichi whom Nayuki was giving the snow bunny was very good. I think KyoAni created the desired emotional impact of Yuuchi recalling the memory. I was impressed.




2. Akiko and strawberry cake

This segment blew my mind. I like the way they blended Yuuchi's realization of the snow-bunny thing with Akiko's accident. The scene of the accident really really shocked me. I cannot put it in words how well done I think the scene was done... you really have to watch the scene to feel the impact. And the way they zoom in on the ruined strawberry cake on the snow was an excellent reminiscence of the ruined yuki-usagi. The cliffhanger was the best I have seen in the series so far.



Plus, the preview was exciting. I especially love the part where Nayuki sat in a corner of her room, blaming herself for Akiko's accident and telling Yuuichi to leave her alone. The next episode is going to become emotionally intense and very interesting.



出ていって、ゆういち。わたしって人も会いたくない。一人でいたいから。

Unfortunately, it seems to me the best of the series is just beginning at the near ending of the anime. I find KyoAni very artistic in delivering emotionally wrenching scenes at the last few episodes of the series. The pattern seems to replicate from their previous production, AIR. The first half was quite calm, and slow paced, but the last 3 episodes made me cry, especially when they showed the link of reincarnation and the inevitable death of Misuzu. The part where Misuzu tethered towards her aunt at the 'goal' end was very very sad.

For the first time, I'm actually looking forward to the next episode of Kanon: Symphony.



All images belong to Memento & Random Curiosity.
Please do visit them for excellent reviews on Kanon.

cross posted in Miharu's Reviews.

～みはる～

Balance of 3

What if you can't graduate even after you have finished all your modules?

Does that sound stupid? Apparently, my university's system is showing me such case IS possible.
Only recently, I decided to try to apply for a Ph.D programme, and the application required me to send a copy of my transcript containing all my exam results. Now, I have completed all my modules last semester, and all was to be done is my thesis project this semester. So logically, I had fulfilled all the required academic units for graduation. I was so confident about the matter, I mean I never fail any module so by right I have no outstanding units to clear, I took it for granted and never double checked my accumulated academic units. When I did check it last week, I almost had a cardiac arrest: there's a balance of 3 academic units.

Naturally, I flipped out.
WHERE DID THE 3 UNITS COME FROM??

Clearly, the 3 units account for one module. I started panicking; it is impossible to register any subject now and I'm already in my FINAL semester. If the balance stays, I cannot graduate!!

When I did calm down, I began tabulating all my points. The math was correct, but the balance should not be there! I tried to check with my cohort, but most were not immediately available. When I finally did get hold of a friend, I realized that the system had failed to display one subject which I passed last semester, which was worth 3 academic units.

I flipped out again.
WHY IS IT NOT THERE?

When I called the university to check, they couldn't give me an immediate answer. After troubleshooting, they told me the result for the subject is pending.

Now, how can the result for the subject I passed last semester be pending NOW? Furthermore, I did not submit any appeal to review my grade. I was extremely pissed off by the fact that they withdraw my results from the system and did not bother to inform me, the examinee, about whatever they wanted to do with it!! What right is there of them to take the liberty of removing the grades and report it as 'pending' without even giving notice to the owner of the grade? My blood boil at the curt reply given, and immediately sent an email back at them demanding for an explaination.

Doesn't that tell you that there is a possibility graduation is not possible despite completing all modules? It must have been on fine prints in the admission letter.

～みはる～

All in 2 weeks

It's been a while since I've posted anything unrelated to lab-life, or any post for that matter in the past few weeks. Lots of things had been happening, and it's not just Chinese New Year.

Two weeks ago while having a barbeque reunion dinner with my housemates and friends in campus, I called home to find out that mum require an immediate surgery. Suffice to say, I was shocked at the urgency and that the reason the surgery commanded immediate action was because her haemoglobin level had dropped to a 'dangerously low' level. However, mum was quite reluctant to undergo the surgery given that her physician failed to give her a clear explaination that would have assisted in her decision making. I was certainly unhappy with that as I felt it was unprofessional of the doctor to not ensure the patient was well-informed before making decision regarding a major surgery. Hence, I decided to take a red eye flight out of Sg and return home to take mum to another physician for a second opinion.

Like I said, I was amidst a barbeque gathering, and thought it would be rude to leave early, given that my housemates had been trying to include me for the gathering (thank you AM, WW! ^.^). Besides, it has been such a long time since I've met up with so many friends and I was dying for a large helping of barbeque-ed marshmallows. And it's not everyday your seniors pay for the barbeque! (thank you 'old people'!! >_<). So we ate and had some good laughs, and by the time I was back in my room, it was way past midnight. I had only 4 hours before I had to leave for the airport, so I decided against sleeping for fear of failing to wake up in time. To kill time, I showered, did laundry, and watched drama. By the time I was to leave for the airport, I felt kinda woozy and lightheaded.

Fortunately for me, I managed to hail a cab quickly, clambered inside, and thought I could catnap while being chauffered to the airport. As I was drifting off, I suddenly felt the cab slowly decelerating. Almost as quickly, the driver stomped on the accelerator abruptly. I blearily opened my eyes, and to my horror the driver himself was DRIFTING OFF as well!!

Now let me tell you, taking a cab driven by someone equally as sleepy as you is certainly no fun ride. My veins nearly popped when I watched the cab veering off the lane and in the direction of the side curb. I didn't know if I should thank my lucky stars when the driver 'regain consciousness' just in time to steer the vehicle back in lane. He was obviously VERY sleepy as he kept rubbing his eyes and ate sweets to keep awake. In the end, I had a terribly unnerving 30-minute journey, all the time praying that we would reach the airport a.s.a.p. before he knocked out cold.

By the time I really did reach the airport, I was very very tired as I couldn't sleep a wink during the terror ride (could you?). It didn't help that the airport was obscenely bright, cheery and full of enthusiastic human all giddy for holiday tours, while I was grumpy and sleepy. After checked in, I grabbed a pretzel and a cup of tea before briskly headed off to the boarding gate. Plonking on a cushioned seat in front of the still-closed gate, I shut my eyes, ignored stares from cheery holidaymakers, tuned out the glaring lights and loud banter, and fell asleep. Fortunately I managed to wake up for boarding, and once inside the plane, I fell asleep even before the plane taxi-ed.

When I touched down, it was already morning, and I immediately accompanied mum to the hospital. We had to go through a series of check ups before the doctor sat us down to discuss about the diagnosis and prognosis. I was much more satisfied with the professionalism of this second doctor, and after hearing out her advice, mum decided to go ahead with the surgery. You see!?! Compassion and soft skills are essential in treating your patients! *fumes at first doctor*

I had to return to Sg immediately as I had not taken any leave from my supervisor. It had been a tiring weekend, and fell asleep again during the flight back. When I did reach my room, it was again way past midnight and all I wanted to do was fall onto bed.

So, the next morning I immediately talked to my supervisor regarding my situation and she was extremely kind to allow me leave until mum recovered considerably. I was very grateful for her kindness, because I was counting on the leave to go home to post-surgery care to mum until she can move around independently. After settling some leftover experiments, I immediately rushed out of lab to catch the next flight back to hometown again. When I called dad on the way out, I was shocked to find out the surgery is already underway. I was shocked because I felt the hospital did not execute the procedure properly and had led to distress to the patient & family, but since the operation is already in the process, I just leave the matters as it is and prepared to travel back home again.

It was already night when I reached home and rushed to the hospital. I then took over dad's shift to look after mum over the night. This is my first experience being a caregiver a.k.a. night-nurse, and I must say it is not easy. It was very difficult trying to keep awake, as I hadn't slept properly at all during the whole weekend. Besides mum was very weak and required attention throughout the night, so it was quite a challenge.

Fortunately the surgery went well without complications, and mum has since gotten better, thanks to all well-wishers. I had taken a long leave to assist mum as long as possible, and had learnt many things along the way. A caregiver's role is not to be taken lightly, and I have come to respect their attentiveness and compassion.

This year's Chinese New Year was comparatively much simpler and quieter than previous years, but just as enjoyable. It was nice to have reunion gatherings because with all work and study overseas, we had hardly have time together as a family. I get to view New Year from my parents' perspective: polishing urns, preparing prayer paraphenalia, vegetable marketing, scaling fishes, hovering over hot stoves, oily pots, endless cooking - basically lots of HARDWORK! The goodness of homemade food and clean altars does not come easy!

Mum had since gotten better from her surgery, but I'm still sticking around home for sometime until she has recuperated enough to return to her routine indepedently. At the moment, I hope my pet E.coli sits in the freezer obediently till I get back to lab working with them!!

Gong Xi Fa Cai!

～みはる～

A Plasmid Fairytale

My experiments lately keep bumping into dead ends, and while searching for information online, I came across this very interesting site. The allegory is very relevant to what I'm currently doing, and I can't help laughing at the personification of the bacterial molecules. I'm sure bioscience students, and those who are not but understand the terms, will find this tale funny XD. Enjoy.


The wise molecular biologist tells us the tale:

Once upon a time, a plasmid met a cell, and they struck up a conversation. Saith the plasmid, "Kind cell, I bring thee a gene for antibiotic resistance, and if you let me in, I shall repay you by showing you how to make an enzyme that might save your life one day." The cell replied with annoyance, "What use hath I for the likes of thee? My food is all gone, I'm freezing my pili off, and you think my biggest problem is antibiotics? Maybe you're just a selfish gene, looking for a cell to make copies of you. Begone!" The plasmid smoothed out his supercoils, dodged a DNase, and tried to think of a suitable reply.

Just then, a massive wave of thermal energy struck them both, and it was all he could do to not lose his footing, standing as he was on the cell membrane. The plasmid trembled, partly to distribute the energy uniformly to his vibrotational degrees of freedom, but mostly at the thought of how painful thermal denaturation might be. Fortunately, it was not coming to that. Not this time. The temperature was high enough to pop a few hydrogen bonds, and his strands were breathing a bit, but he was covalently closed after all. He could handle it!

The cell was not so lucky, however. His membrane, which had been reasonably firm to this point, began to swirl and form vortices, and lost its smooth surface. The cell had gotten used to the cold temperatures by boosting the fraction of short chain and unsaturated fatty acids in his membrane. While it gave the membrane a good consistency in the icy cold, it was completely wrong for this new, higher temperature. The van der Waals interactions weren't strong enough to maintain cohesion, and errant lipids were now leaping through the bilayer like divers at a mosh pit. Huge patches of the membrane were involuting, bringing massive gulps of medium inside. The cell was just seconds away from a complete membrane breach!

Then, as quickly as it had started, the problem was over. The buzz of thermal energy was drawn away by some unseen entropic sink, and dissipated. The membrane returned to its glassy-smooth state. The plasmid was now inside the cell, having been carried through the membrane in its moment of weakness, but the cell was too preoccupied to notice the uninvited guest. The cell was using his last remaining energy to bail out the excess liquid, and set his ion gradients to rights. When he thought he could do no more, that his determined course would be senescence and death, a warm flow of fresh medium restored his spirits.

"Ah! Tryptone, and yeast extract", he said as he gratefully derepressed half a dozen operons. His ribosomes got right to work, making the enzymes that would help to catabolize the new food source and restore the structure of the cell to full health. He almost exclaimed, "I thought Darwin had me for sure that time!" but being a cell with little memory he had already forgotten the privations of a few minutes earlier. Thirty minutes passed, and he found himself cast onto a wide surface that was warm and rich in nutrients.

"Not bad", he said. I could live like this for generations!" But then, the horrible sounds of dying cells reached him. All around him, his brethren were being killed by an unseen attacker. An antibiotic was there, and was destroying the entire population. Oh the humanity! He braced himself for death, but then ... nothing happened. He was alive!

"Remember me?", said the plasmid. "I told you that I might save your life one day, and now it has come to pass." The plasmid, which had gone unnoticed since the thermal catastrophe, had been copied several times and transcribed by the cellular machinery. It had provided a gene that encoded an enzyme that destroyed the antibiotic before it even got into the cell. Although more antibiotic was diffusing into the neighborhood, the enzyme was on the job, and prevented it from doing any damage.

"Yes, you were right", said the cell. "I am grateful that you transformed me, and now that I have a logical explanation for my good health, I don't have to develop survivor guilt either. Stick with me, plasmid, and I'll make sure you are provided with a high copy number."

The generations passed, and the cell divided many times. Each time there was fission, the two daughter cells received an inheritance of plasmid copies. There was widespread prosperity.

I would say "They lived happily ever after", but sadly that is not the end of the story. The daughter cells grew into a prodigious colony, and soon had destroyed so much of the antibiotic in the immediate vicinity that the real danger had passed. Some cells that had not perished in the original attack even managed to survive and grow nearby - little "satellite colonies" seeking refuge from the high levels of antibiotic elsewhere.

Then the cells in the big colony became lazy. The plasmid was not replicated to the same high copy number - it no longer served the interests of the cell to do so. Sometimes, daughter cells did not inherit even a single copy of the plasmid! As the colony aged and grew, the proportion of cells that carried the plasmid became less and less.

The colony actually believed itself to be quite progressive on this point. Some of the more strident cells even argued: "Why should young cells be forced to make enzymes that they don't need?" They said "We want to evolve higher order characteristics, not merely regurgitate the knowledge that served our great-great-great grandcell!" Before long it was unfashionable to carry the plasmid, though a few still did, but the colony grew faster without the added responsibility of the added synthesis.

One day, a toothpick scraped the colony from a plate and carried it high into the sky. "At last," the progressive cells thought, "we are entering a bold new era in which we are going to be able to realize our true genetic potential!" The toothpick was dropped into a tube of fresh medium. The fresh medium had fresh antibiotic.

What happened next is almost too horrible to tell. The cells that still carried the plasmid lived of course. They produced the enzyme that destroyed the antibiotic. The cells that had not inherited the plasmid were unworried at first, because they thought that a protective shield of enzyme could be built by the others, but being in liquid medium there was no hope for them. The antibiotic was not limited by diffusion, as it had been on the plate, and they were soon overcome.

The few wise cells that had not lost their plasmid went on to eternal storage in glycerol stocks, and were written about in books and famous journals. They were grown in huge 10,000 liter fermentation tanks, and provided with the very richest medium that had ever been made!

The end.


～みはる～

Colony, where art thou?

Creating a molecular library is no easy feat. I almost landed myself in a phenomenal agar plating exercise.

Finally after a month of preparatory work, I was ready for my first trial run of molecular library construction. I was pretty optimistic, because I had everything carefully planned, right down to the number of eppendorf tubes that I had to use. I even took the pain to read several different protocols and scrutinize my experiment planning from all (perceived) possible angles. The only thing I did not do was read the directions of how to use the commercial competent cells properly. When I actually did, I came out with this mind-blowing numbers:

To make 1 library, I need:

• 400 eppendorf tubes for cell transformation
• 2000 agar plates for spreading of library culture

Because I have 7 library combinations, I need:

• 2800 eppendorf tubes for cell transformation
• 14000 agar plates for spreading of library culture

Essentially, I have to transform 2800 tubes of cells and spread 14000 agar plates in one go.
Say I take 1 minute to spread one agar plate, it would take me 233 hours, which is approximately 24/7 for 10 days, to finish spreading all the plates.

-_-'''

That is if I sit at the biosafety cabinet (BSC) for 10 days straight without leaving the BSC even for a split second.

My eyes nearly popped out when I saw the ridiculous figures. Forget the BSC-thing, even if I put all the incubators in the lab together, there isn't sufficient place to incubate 14000 plates!!

Now don't even entertain the thought of me spreading 14000 plates. Anybody knows that is quite impossible to be done. However, my supervisor, mentor, colleagues and myself had a good laugh at the scale of the experiment. We then worked out an alternative strategy to do the experiment, MINUS 14000 plates. *phew...*

So, after tweaking the protocol a little, I run the trial experiment at a manageable scale. I followed the recommended protocol strictly with hopes to generate as many colonies as promised by the company protocol. When I put in the plates in for overnight incubation, I was quite optimistic and very hopeful to see a lawn of colonies when I return the next day. I mean, afterall, they did say I'm supposed to obtain millions of colonies!

I waited and waited and waited till the hour came, ran to the incubator excitedly to take out my plates. どきどき。My heartbeat increased as I anticipated the lawn of E.coli on my plates. Chuckling gleefully, I reached for the plates and turned them over for a good look....... NO COLONY! I reached for another stack of plates..... ALSO NO COLONY! My heart sank as I reached for the final 3 stacks.... SPARSE COLONIES!

"Oh nooooo, what had happened to my cells?? I have no library??!! Even with using the most competent cells?? Dieeee......," - was all that was running in my head.

I supposed research isn't without experimental failures. Now I'm tweaking my strategies again and hopefully my next round of trial will bear fruit... or rather, fat juicy colonies.

Don't tell me I really have to spread 14000 plates?....

～みはる～

バカなことばかり

You know, I think I may have fried my brain cells or something. Lately, I've been doing many stupid things.

It must have been my incompetence, as I seem to be multitasking most of the time. Take last Friday for example, I went to lab very early to do vector control and to run (the umpteenth) genomic DNA extraction at the same time. Kind of stupid, really, to run two experiments together, but I was greedy and didn't want to waste a second of the waiting time. To add stress to it, I had to finish transforming ligated DNA and extracting gDNA both by lunchtime as I had a career workshop after lunch. Now, anyone experienced will laugh and tell you it is impossible to complete so much work in half the day. But I was greedy, laid out the materials on two opposing benches and spent the entire time going back and forth from one bench to another. On one bench the centrifuge is done spinning and keeps beeping, on the another bench the digested mixtures are completed awaiting for gel electrophoresis. Gyaa.... what a dizzying exercise.

In the end, I couldn't finish the experiments in time and was late for my workshop. My mentor had kindly run my DNA extraction gel electrophoresis for me, as I skipped lunch to rush to the workshop. After the workshop, my mentor told me my DNAs were weird. The extraction yield was very good, but there were EXTRA bands. I was like, "nooo.... my whole afternoon is wasted again!!" I quickly went back to lab to troubleshoot and re-run the gel using one of the 16 samples, and found no extra bands O.O The DNA must be playing hide-and-seek with me.

However, I still could not leave as my transformation part of the experiment was not completed and I had to wait like an hour for the bacteria to grow before I can spread them on antibiotics plates for overnight incubation. When I went to the cold room to get plates, I realized there were NO MORE PLATES!!!! The antibiotic plates I wanted were not in stock!! Talk about bad luck! T_T What were left were a stack of plates labeled "Not-so-well-made-Amp-plates".

-_-''' I do not know what the person who made the plates meant.

I had no choice but to use them anyway, and because these plates were 'defective', I had to plate on another type of plate which were good quality, but had slightly different nutrient composition. Meaning, I had to spread TWO sets of plates. Now, you might think, another extra set only mah, what's the big deal. It means I had to spread 13 samples on 52 plates. And that time was already 930pm.

By the time the bacteria had happily grown to the state I wanted them to, I was the only person left in the level again. T_T I don't really enjoy OT, because the quiet lab can be quite creepy! I think it must have really just been me, but I keep hearing sounds. However, thanks to that, I have learnt some plating-acrobatics, as I could now spread plates without even looking at the plates (half the time my eyes were averted nervously to see if any vampire, uh I meant person, sneaked up behind me).

Then yesterday, I went back to lab to take out the incubated plates and do genomic DNA extraction A.G.A.I.N. (when can I get enough DNA??) Fortunately, my vector control went well; meaning I can proceed to actually start making the molecular library. After finishing my DNA extraction, I run all my 32 samples on gel to check the DNA concentration. I had purposely made brand new gel from clean TBE liquid and ultra-grade agarose, so I was confident my gel quality was good. Furthermore, I had performed the extraction using the conditions I optimized last week, so I was quite sure my yield would be pretty good.

So, I loaded my samples, covered the tanks, and plug in the cables. As I was connecting the cables, I noticed the tank orientation was upside-down. "But it's okay, as long as I plug the electric poles right," I thought. After running the gel for 45mins, I took them out of the tank for EB staining. As I did, I thought something was weird. Usually, the blue dye from the samples can be seen at the end of the gel after electrophoresis, but I didn't see them on my gel. "Maybe my gel's a bit too short, but it's okay, since the bands are not very low," I had thought. So, without further thoughts, I put both pieces of gel in for staining... and waited 25mins for viewing.

After staining was done, I took the gels to view under UV. What I saw on the computer nearly made my heart stopped beating.

NO BANDS.

When I viewed one gel which had two rows of wells, I noticed something odd: the bands from the lower row appeared on the upper row gel. At first I thought I placed the gel upside-down on the platform. But it was quite impossible. because the ladder was on the correct side!

Then, it dawn upon me.

I had ran the gel BACKWARDS.

The electrical poles were right, but the gel was oriented upside-down. Thus, all the DNA from the upper row were run into the tank liquid. How stupid can it get?? I didn't know whether to laugh or cry when I saw the gel photo. I mean, WHO RUNS GEL BACKWARDS??

I gave up for that day. Maybe it's because my concentration level had hit low and I'm beginning to do very unsensible things.

Lesson is.... always keep the tank on the right orientation.

～みはる～

Mad rush

The last three days had been, like, experiment-mad-rush for me. I've been clocking in very early and staying in the lab till very late designing and executing experiments to troubleshoot my thus far negative results.

On Thursday, I stayed late to extract the bloody genomic DNA which turned up with terribly poor yield. The entire day of lysis - centrifuge - lysis - centrifuge - wash - centrifuge - wash - centrifuge - elute - centrifuge - elute AGAIN had gone to waste as proven by the blank gel photo. I darn hate it when I get blank gel photos. It didn't help that my gel broke into half and I wasted the time to piece it together, when all I actually had was no DNA.

Yesterday, I left the lab only when midnight struck. Seriously, I might consider sleeping in the lab soon. The whole day and night was devoted to making.... you've guessed it, competent cells. Chemically competent E.coli in the day, electro-competent E.coli in the night. By nine, most of my colleagues and FYP-mates had left the lab, and then I realized I was the only living being (apart from all the bacteria in the freezer) in the entire level. Heck, it was possible I was the only being left in the building. The thought creeped me out, especially when I had to work at the biosafety cabinet (BSC). While aliquoting the stupid (hopefully) competent cells into small vials in the BSC, my back was facing the lab entrance, and not being able to see anyone who might come thru the lab entrance made me very nervous. All the while aliquoting, I thought I heard the hinge of the lab door creak. Obviously there was no one around, but I kept turning around in paranoia and the pipette in my hand started to tremble so much I couldn't aliquot properly. After several minutes, I was no longer creep out but very annoyed because the volume of cells made didn't seem to reduce no matter how much I transfer into the smaller vials!!!! I was already very very tired and wanted to go back to my bed, but the bottle of cells remained still as much!! When I finally did finish with the cells and cleaning up, it was already close to 11pm. But then, I did something very stupid.

After I left the school building, I realized I forgot to transfer the expensive rotor back into my lab from the common room. My lethargy was getting into my head, and I thought, "I'm too tired to walk back to lab. I'll come back earlier tomorrow to keep it." However, after walking some 800m away later, a wave of worry washed over me as I begun to imagine unauthorized users spoiling/stealing the rotor. There is absolutely no way I could pay for that bloody expensive rotor. Besides, if the rotor is spoilt, I can kiss my FYP goodbye. It was a storm in my head as the Angel and Devil debated whether I should continue walking back to my room or to return to the lab. Finally, Angel won and I trudged back to lab. Stupid rotor! By the time I really did reached my room, it was already 12midnight! If I had turned back the first time, I needn't wasted my already-draining energy and precious time!

I thought I could then just sleep after such a long and tiring day. But no no. I had another experiment to design for the next day and had much thinking to do. I didn't really have the mental energy to think, but I kept tossing and turning in bed, until I gave up sleeping and got up to design the experiment at 3am.

I didn't had it easy today either. Had to wake up early to go to lab again to troubleshoot the genomic DNA extraction and transform the (hopefully) competent E.coli cells. I had thought I could finish before dinner time, but like always, E.coli dictates my schedule, not vice versa. In the end, I get to leave the lab only at 9pm... and I didn't even get to run gel to check the gDNA!!

Tomorrow's another half day (I hope) in lab again... and this time, I'm praying all my results are positive. If they aren't, I declare war against E.coli.

～みはる～

Demoralized

I'm feeling so so demoralized.

Already I had no competent cells after one week of making it, and today's genomic DNA extraction failed miserably. What is very demoralizing was I spent the whole day painstakingly extract the stupid bacterial DNA and after running the samples on electrophoresis, the bands were BARELY visible against the freakingly bright DNA ladder. ONE WHOLE DAY spent and NO BANDS! All my 16 DNA preps were gone to waste.

This is, like, a week of experimental misfortune.
Nothing is going right.
No positive results.
How demoralizing.

～みはる～

E.coli-tachi, たのむよ！

I've made more than 200 tubes of E.coli cells, and yet, NONE of them are competent!!! T_T

You see, I'm creating a bacterial genomic library for my FYP and as with all standard protocols, I need competent hosts that can efficiently carry the genome. So, I had to make a fresh batch of competent E.coli cells for my purpose. After spending a week making 2 batches of cells amounting to more than 200 vials, NONE of them passed the efficiency test. And now my whole molecular library construction is brought to a halt because I couldn't generate the right host!! To make the matter worst, I don't know why! And to top it off, the procedure is taking too freaking long!!! The incubation period is like 16 hours, and I have twice come out with barely a handful of E.coli colonies on the plates. T_T

To the E.coli on the 24 plates sitting in the 37 degrees incubator right now, please...G.R.O.W.
たのむよ！

～みはる～

It's the MODEM

Last weekend, I suffered the Internet-withdrawal-symptom.

My Internet connection was active but loading of websites all timed out during operation. Meaning, I couldn't connect to the world for an entire day!!! Losing this connection is like a drug addict losing his daily fix!! I could not do anything the whole day, but frantically loading and re-loading the pages, trying to *unsuccessfully* fix the problem. In the end, out of desperation, I sms-ed a couple of friends to ask what could be done for me to get my Internet dose. Guess what? The solution was very simple. So simple that I could not believe I wasted more than 24 hrs without Internet just because of this.

Switch off, and restart the modem.

I learnt my lesson.

This scare tells me that my lifestyle has evolved such that my daily life revolves around the Internet service. I NEED the net for animes, dramas, news, blogging, researching and most importantly connecting with people. Without it, my world would really fall apart. I mean, can you imagine not getting the latest episode of Death Note???!!!!!!

～みはる～

研究の生活

最近は、すごく忙しかった！！
卒業プロジェックトもう始まったし、毎日朝から晩まで　研究室で実験をするばかり、もう暇な時間がなかなかありませんでした！
でも、どんな忙しくても　やることがあるなら　日々ももっと楽しいと思います。＾＿＾

Since my return to campus, I have been so tied up with my FYP that I spent more than half my day in the lab! It's like 9am reach lab, 9pm leave lab... for at least 5 days a week. Although such schedule is very tiring and leaves me no time for dramas/animes at night, I find that having things to occupy me the entire day quite fulfilling and gives me a purpose to look forward to each day. Well... my current purpose is to get the bacteria to like me so that I can complete my FYP >_<

じゃ。。もしひまだったら　今度また何か書くね。

～みはる～

あけましておめでとう

Happy New Year!
May you be blessed with love and good life.

あけましておめでとうございます！
いろいろありがとうございました。
今年もよろしくお願いします。

～みはる～