He's done it again, Ronaldo boy. For 2nd year in succession, Cristiano Ronaldo is named PFA Player of The Year award. He won the award after beating his close rival Fabregas and Adebayor (Arsenal), Gerrard and Torres (Liverpool) and David James (Portsmouth). This is the 2nd time a player winning the award in 2 successive years after Henry did in 2003-2004. His form has been irrepressible for most parts of this season, scoring 38 goals in all competition, including one of the most creative, fancy goal in their 4-0 victory against Aston Villa. Now Man Utd is still on course for a double (Premier League and Champions League), should he be able to keep his form high in the rest of the remaining matches, then the EPL title will be theirs again, and the UCL trophy will return to Old Trafford for the 3rd times.
Talk about the award, he humbly (lol) gives tribute to his teammate for his achievement, and as usual, his typical comment as seen in many of his post-match interview. Quoting him; "I feel very happy. When you work all season to play good, to do something for the team, to do your contribution for the team and then at the end of the season the PFA give you this award, it is a great moment, it is an honour, a pleasure. It is a great motivation to carry on like that, to work more and get better. I feel very good but it is not just my award, my team-mates have helped me a lot this season. They give me good passes to score goals - and I score - but it is a good moment for me".
Microbiology Mid-sem test sample: You have cloned into E. coli, a gene that may encode resistance to the antibiotic chloramphenicol. This conclusion is based on the fact that the DNA sequence of this gene has similarity to that of other choramphenicol resistance genes. Explain how you could use gene disruption analysis to determine whether the gene does indeed confer resistance to chloramphenicol in the strain of bacteria from which you cloned it.
sample answer:
(a) Transposon method: Assume the original host is resistant to chloramphenicol. A suicide plasmid carrying a transposon would be transferred cells of the host bacterium by transformation or conjugation from an E. coli culture. Let us assume that the transposon also carried a tetracycline resistance gene. The transformed celles (or the tranconjugants) would then be plated on growth medium containing tetracycline. For the tetracycline gene to survive, the transposon must hop out of the suicide plasmid into othe host DNA. Since this random event will occur in many millions of cells, some cells are likely to contain transposon insertions in the putative Cm gene. We can select for cells containing insertions in the Cm gene by plating all tetracycline resistant cultures on duplicate plates containing both tetracycline plus chloramphenicol. Cultures that do not grow in the presence of Chloramphenicol must contain insertions in DNA involved with Cm resistance. To confirm that the transposon had inserted into the Cm gene, we could use PCR technology to sequence DNA flanking the transposon to determine whether it is identical to that obtained previously (see question).
(b) Single crossover (or cointegrate method). Assume the original host is resistant to chloramphenicol. A suicide plasmid containing part of the cloned gene could be introduced into the original host by transformation or conjugation. Assume the plasmid also has a tetracycline resistance gene for selection purposes. When the transconjugants or transformants are grown on medium containing tetracycline, the only way the plasmid DNA can survive is by integrating into the host DNA. This will be facilitated by recombination between the part copy of the Cm gene and the wild type copy in the host. This event will result in integration of the entire plasmid into the host DNA, thereby creating an insertional mutation. Tetracycline resistant cultures would then be plated on medium containing tetracycline + chloramphenicol. Lack of growth would confirm that the Cm gene had been insertionally inactivated. This could be confirmed by PCR amplification of DNA encompassing the point of insertion of the plasmid DNA followed by sequencing of the host DNA flanking the insertion. If the sequence data was identical to that obtained previously (see question) then you could say that the gene encoded Chloramphenicol resistance.
(c) Allelic replacement. Assume the original host is resistant to chloramphenicol. The cloned copy of the gene would first be insertionally inactivated in E. coli by cloning a copy of another gene eg tetracycline resistance gene into the Cm gene. The construct would then be cloned into a suicide plasmid (carrying an ampicillin resistance gene for selection of the plasmid) and transferred to the host cells by conjugation or transformation. Transformants or transconjugants would be selected by plating on tetracycline medium. Tetracycline resistant colonies would then be plated separately on growth media containing tetracycline + chloramphenicol and Tetracycline + ampicillin. If a double crossover recombination event had resulted in exchange of the mutated copy of the Cm gene into the host, then no growth should be observed. This would be evidence that the Cm gene encoded chloramphenicol resistance. This could be confirmed by sequencing across the junction between the tetracycline gene and flanking DNA. Any cultures that were able to grow on Tet + ampicillin would represent cointegrates formed by single crossover recombination events. These should also be chloramphencicol sensitive.
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I was liek....omgbbq this is what I'm supposed to answer in the test??? I can hardly remember reading those things in the lecture notes, probably because the lecturer is so boring, or I actually miss the lectures in week 5 when I was infected by the virus... revision, revision...uh 20% of final marks for Microbiology, seems like I need to cram like Konata :3
OMG I forgot that yesterday this blog reached it's first anniversary!!! お誕生日おめでとう!!!
Hm...still remember the time around those period; I'm sitting on by bed, a little frustrated after Manchester United only manage to get a draw in one of last season EPL match. Out of boredom, I lay down with my laptop in front of me, and decided to make this blog, even though I rarely has the motivation to write something... and recently shit things happened, I can't seem to change the template of this blog. Error messages always appeared whenever I try to upload my new template T_T p/s: mid-sem holiday progress: *Physiology lecture notes - finished! *MBS Prac B/NCBI worksheet - 50% finished *MBS Prac A Report - 50% finished *Biochem lectures - the voice recording is sooo unclear =( *Microb test preparation - 0%
totallnewbie shows a creative way to manipulate waypoints and the special feature of WTF mode. After all those "The Art of Mindgames" series, he came out with a showboating video, which imo is nice to see. Starts with Puck, alternating Orb and Rift, followed by Coil (and still with Rift in between). Then Invoker, alternating Meteor and Blast. And finally Antimage with ninja-style blink to dodge the attack of the Fountain. Notice that he does not take a single hit from the attack.
This match is among the highlight of the first round in ESL Group B. On the spot, most would be amazed with the lineup that MYM use: Pudge and Techies. Okay, maybe Pudge is not that weird, but Techies? The freakin Squee&Spleen the Goblin Techies? Yeah it may look a bit offensive and disrespecful to the other team, but if you look again, it may just be another revolutionary move by MYM lol. We've seen some unusual picks before, some manages to give them victory (eg: XsK using Rikimaru in some of their matches) while others failed (eg: Sweden uses Spiritbreaker in their Prime Nation match against Kazakhstan); and in this match, Techies is virtually nonexistent and almost always make the match 4 v 5 in every situation. Not because Hanni plays that bad (well if you compare with Merlini' techies, it does look bad though XD), the hero itself is generally does not suit with the current metagame. However, this handicap does not stop MYM from pulling a victory XDD
Merlini missing from the lineup certainly makes some fans disappointed, but Maelk, Fear, Mania and Lavent play really well with good teamwork to make up for the weakness in Techies.
p/s: one hilarious moment in the match happened in around 34th minute mark when Techies try to suicide on Spectre with his teammate (Tiny and Lina iirc) closeby, unfortunately Spectre manage to disperse the suicide, hurting Tiny A LOT, leaving Spectre an easy job to finish him off :3 -the moral of the story: don't try to suicide on Spectre when your allies are nearby XP
Plot Summary:Craft Lawrence, a 25 year old peddler travelling from town to town selling and buying various things to make a living during a period much like Europe in the Middle Ages. One night when stopped at the town of Pasroe, he finds in his wagon a 250 year old pagan wolf deity girl named Holo. She appears to be that of a 15 year old girl, except for a wolf tail and ears. She introduces herself as the town's goddess of harvest who has kept it blessed with good harvests of wheat for many years. Despite having the responsibility to watch over the town, she wants to go back to her homeland in the north called Yoitsu, she believes the people have already forsaken her anyway and that she has kept her promise to maintain the good harvests. She manages to bargain her way out of the village by making a deal with Lawrence to take her with him. As they travel, her wisdom helps increase his profits, but at the same time, her true nature draws unwanted attention from the church.
Spice and Wolf (Ookami to Kyoshinryo) is an anime adaptation based on award-winning visual novel with the same title by Isuna Hasekura and was scheduled as part of Winter 2007/2008 anime season. It concludes on 25th March 2008 with 13 episodes; 12 episodes for actual TV series and 1 episode of OVA which will be released later.
Spice and Wolf is by far one of the best shows I’ve watched in 2007/2008. One thing I like about this series is the theme. It’s quite rare to see an anime using economy, merchant and trading as a theme, yet manage to give fairly good impression toward the audience. This theme may be encompassed in a possibly generic storyline - a young merchant named Lawrence travels across the cities to trade items and later meet a wolf-deity girl named Horo, and together they start the journey to the north, the place where Horo belong; however what makes it interesting is the series of events happened along their journey from town to town. It’s all about profit!
Story: This area gives me a mixed bag of feeling, but overall I’m more on the positive side. It’s always a pleasure watching the banter of Lawrence and Horo in almost every episode, having some nice conversations and critical analysis on the economic stuff then ran into. Horo is undoubtedly the center of attraction in almost every episode, showing a strong characteristic and pride (that is, in her tail). Her loneliness which was depicted in several occasion were outweighed by her bright, adorable and cheerful characteristic shown whenever she spend her time with Lawrence. The voice Actor (Ami Koshimizu) did a splendid job to give Horo an outstanding characteristic. It appears that Horo voice sounds more erotic and appealing that what the subs appear, so it’ll be more enjoyable if I fully understand what the character says. As for the economic stuff, I wish they could give a better and creative means to convey them to the audience, like what Hikaru no Go did with Go board game. Sometimes the way they tell the details about the trade, the silver content and such are very linear and boring. Well at least to some extent Horo did a good job to explain certain thing in a simpler way to solve the puzzles. And lastly, it seems to me that due to the short length of this anime, sometimes the story looks compressed that some details are failed to be covered. Cloe for example, I think she plays very little role in the overall story of Spice&Wolf. She appears early in the series as Lawrence apprentice, and later with a link to the church, become a threat to Horo. But that’s all she had to offer, despite being an original anime character (in place of someone else, according to the novel reader) she totally miss the climax of this series. I was hoping she could give a bit more impact to the overall story, especially for the final 3 episodes where the real conflict arises.
While the animation quality is not something to be proud of particularly in character animation area, the way they describe the Medieval Europe setting is very impressive, with in depth detail of building structures and environment. Character design is fairly simple, reminding me a lot to Princess Resurrection. The main characters (Horo, Lawrence, Nora and Cloe) somewhat gain the most out of it, making most of the side characters a bit disappointment with many of them lacks variation and look unappealing.
As for the music selection, I have to say that the BGM used suits very well with the scenarios and theme of the series, especially because they’re using actual instrument rather than synthesizer. While many of the tracks are not really fit to be put on your music playlist, they certainly sounds unique to Spice&Wolf and likely more recognizable, just like the unique BGM tracks in titles like Rozen Maiden, Mai Hime and Hayate no Gotoku. The opening song, Tabii no Tachuu by Kiyoura Natsumi is one of the sweetest OP I’ve listened to, and while I’m not that fond with the ending song (Ringo Biyori by Rocky Chack due to the “Engrish-ness” thus made me hard to follow the song without actually reading the lyric. But somehow I grew attached to the song after listening to the full version of the song, although if you read carefully to the lyric, it kind of does not make much sense to the story.
Overall Spice and Wolf is very enjoyable show to watch. Sure it would be boring if you didn’t enjoy the economic stuff, but for me the presence of Horo is enough to make this show interesting in its own way. In my anime dictionary, I would normally give this one a 10 out of 10 rating, but considering some weaknesses that this series posses, I would give this 8.5/10. I hope with that kind of ending shown in the final episode, it will open an option for second season, which I would be more than happy if it really comes out.
Artist: Mizuki Nana Title: Secret Ambition Magical Girl Lyrical Nanoha StrikerS OP Single
Romaji
Mune ni yadoru atsuki suisei wa hajimari no kodou he Furueru yubi nigirishimete shizuka ni negai wo kometa Usukumatte nigerarenai kako kara tobi dashitaiyo
Tatoe kiesou na wazuka na hikari datte Oikaketai ano hoshi no you ni Subete ukeire sou, dokomademo takaku
Ginga wo mau tenshi no sasayaki Tashikana kioku wo tadotte Korekara kitto umareteku shinjitsu he no tobira Donna tsumetai kurayami ni shibararete itemo Boku wa shiritai kara kesshite tomaranai
"Nakusumono wa nanimonai" to chiisana kata wo dakishime Kizutsuketeta itakutemo tsurakutemo modoranai kara
Ima wo kaetakute muchuude kakeru keteta Demo, kitsukeba daijyoubu dayo Itsudemo kimi ga tonaride waratteta
Sora ni ochita tsurugi no shizuku ni mihateru omoi wo kasanete Soba ni aru yasashii ondo ni mada furerarezuita Sono yokogao ni hontou wa kokoro hodo kitai Boku wa shitteru kara kesshite hanarenai
Mamoritai to hajimete kanjita Ashita heto tsuzuku kono basho Mai agare bokutachi no yume hitori janai I don't forget
Tenshi no sasayaki tashi kana kioku wo tadotte Korekara kitto umareteku shinjitsu he no tobira Donna tsumetai kurayami ni shibararete itemo Boku wa shiritai kara kesshite tomaranai
The hot comet dwelling within my heart beats toward a new beginning My shaking fingers, grasping upon the silent wish Cowering, not wanting to flee from the inescapable past I want to do it all at once
Even if it is merely a fading, faint light I want to chase after it, just like that star Accepting everything, no matter how high it is
Dancing throughout the galaxy, diamond dust is the angel's whispers Perusing these certain memories The door to truth which will definitely be born now No matter what kind of cold darkness binds me Because I want to know, even death won't stop me
"I lost something, something and nothing" hugging a small shoulder Because my feelings hurt a lot, but I could not turn my hand back without difficulty
I begin to change now However, I want to feel "safe" You will forever be my neighbor, smiling
The sky fell onto the sword, dripping down[1] Once more, the thoughts are incomplete Besides, there was tender warmth when I was touched There was truth in that profile, I wanted you to unwrap my heart
I want to hold onto that first impression Tomorrow, I will continue to be exhausted here Our dream will fly high. it's not one person I don't forget
The angels talk of diamond dust I wonder if you follow a certain memory The door to truth that will definitely be born now No matter what kind of cold darkness binds me Because I want to know, even death won't stop me
Translation Notes:
[1] In the kanji version, there was an extra surugi ("sword") in this line. Since the romanji version and the karaoke version don't have this, I omitted it. The extra words in "-" like -surugi- and -tenshi no sasayaki- in the kanji lyrics are how to pronounce the previous word(s), 月剣 and Diamond dust respectively, much like the hiragana/katakana text over kanji. That being the case, these words are not repeated in singing, nor do you hear Diamond dust in English anywhere in the song. self-translated
It's kinda irony as I never have any interest in any Nanoha series, yet I got attached to this song for so long. I've tried watching the first season (Mahou Shoujo Lyrical Nanoha) but I simply can't digest mahou shoujo series, including this one. Nevertheless they have some of the best singles, especially those by Mizuki Nana.
One spot was left empty in Group B of the ESL Major Series Season II DotA league.
A lot of people have expected it to be filled by Virtus.Pro, which is one of the most successful DotA clans to date.
The replacement team however, is far from anyone has expected. XcN, the best Indonesian team, will be the one filling the empty Group B spot.
Antony "therogue" Seedhouse, the head admin of the tournament, mentioned that he has invited a lot of teams such as VP to join, but either said no or did not reply to the invitation.
XcN was chosen because they have shown the initiative to be part of the tournament. XcN wanted to play the Asian qualifiers, however that section of the Major Series suffered no sign-ups.
XcN is the only Asian team in this tournament. They will be facing teams such as Meet Your Makers, The Elder Gods and Coup de Grace in their group.
I was liek, lol what an unexpected filling. Being the only Asian side in the tournament, one very expected problem that this team will experience is the difficulty to look for Neutral Host, as well as lagging problem. Kingsurf from Malaysia has already shown how difficult it is to find good neutral host when playing against European team, and often they plays in lagging condition. So XcN may experience even greater problem, since Indonesia internet connection is generally worser than Malaysia. Anyway, GLHF to XcN in this tournament =)
rumor has it, whether it's true or not the jersey looks cool anyway (albeit a bit plain as always), even though it looks a bit like USA international jersey.
Sometimes it's pretty fun to play, just to kill some times. I haven'tfully explore the mechanic of this game, so far I only manage to get around 2000+m. I've seen people scored as high as 9000+m though.
For unknown reason Anime Saimoe is unlikely to happen for 2008, but in turn, we have a league format Saimoe taking place of the traditional Saimoe. Dubbed as International Saimoe League, this is where 64 popular characters who ranked the highest in previous Japanese and Korean Saimoe will battle each other in league format. The top 16 characters in the league will qualify for the playoff with double elimination format.
2 weeks mid-semester break has just began, while it may be a bit leisure times for me to relax, there's still a lot of things I need to cover during this break. First, Microbiology lectures. The current lecturer (Dr. Connor Thomas) makes me yawn so much that I "have" to skip his lecture :3 The number of his lectures that I attend are countable (once a week, sometimes I skip the whole lecture in the week). So a lot of lecture notes need to be revised, and to top it all, I got an important test coming right after the break, cumulating 20% of total marks for Microbiology subject. Then Physiology. I like Prof. Andrea Yool lecture, but we as Biotech student were not able to attend Friday lecture due to class clash with Biochem tute, so one lecture is guaranteed to be missed every week. And in week 5 I had a terrible virus infection for the whole week, so I missed all her lecture for that week, and 2 lectures for week 6. Luckily I barely made it to submit Literature Review for Practical last Tuesday, and the tutorial is easy as ever. And finally Biochem. I loooooove Lynn Roger lectures, so I never missed any of her lecture (except in week 5 when I was sick). I've done with the test on Friday, as well as the online test. So nothing much to worry about this subject. So what left is MBS Prac A and B. There's a whole load of reports need to be submitted after the break, particularly the Bacterial Technique, Recombinant DNA and Enzyme Kinetics. While I haven't finished analyzing the uv gel photo for both DNA ligation and recombination. Same goes for Prac B, a lot of works need to be done on NCBI database and Jalview.
So here I am, while having this free time, writing this piece of shit. I won't be able to play on GG-Client for the past several weeks because my connection become unstable whenever I play and disconnected halfway through. So frustrating... So for the time being, I'm Cabal-ing :3
p/s: lately I'm addicted with a lot of songs from Mizuki Nana :3
We'll find the path to Alsatia We'll lose if we don't fight now
No one can play in the sculpture garden Petrified faces are watching in the dark Assimilate yourself in this mystical world That's all you have to do
Day after day, you got tragic situation Night after night, you tried to break the wall Fortune is calling you in this mystical world You'll find a way
Cloud in the mists of time You must take your aim and play your game Emotional neglect is the price that you must pay to your desire It's Alsatia You just believe in the lie to forget the awful truth You know the disturbance never ends
There is no reason to take this process Resentment will change into sweet desire You'll run for your life in this mystical world Beware of the night
Crawl in the mists of time You must face the wall of the hate Eternal vigilance is the price that you must pay to your desire It's Alsatia You just go against the law, fall into a delusion You know the distrubance never ends
Cloud in the mists of time You must take your aim and play your game Emotional neglect is the price that you must pay to your desire It's Alsatia You just go against the law, fall into a delusion You know the distrubance never ends It's Alsatia You just believe in the lie to forget the awful truth You know the disturbance never ends
We'll find the path to Alsatia We'll lose if we don't fight now