Fuck Had a terrible morning all bcos of a freakishly fucking teacher. Ppl think so long to draw de diagram then you make them a fool infront of other ppl. Wad, "where got *** diagram like that draw one? Dunt give me this nonsense!" Well I tell you, you are fucking nonsense. I took the initiative to ask you a question, you ignored me, and degraded me and my work, somemore that someclassmate still say yalor. What the fuck man.
Liddat still not enough, still after explaining de question must tell de "mistake" in front of the class. I must have looked like a complete fool in some ppl's eyes. Fuck
And wad's more? You ignored my hardwork, you ignored my question after a look at my diagram. And my answer was right, my concept was right, the diagram's divisions were right. But wad did you do? Not a positive remark abt them.
I know that you are so called "senior" and going through your menopause but don't let that fucking excuse affect your teaching! And do you think i give a shit abt excuses?
stop looking down
brightness will break through, and SHINE @ 5:22 AM
Friday, April 18, 2008
I just realized during the short span of 3 days i missed out alot! well actually more than 3 days... Things like robin's bday, keith's bday and more... well there is more to it than wad is seen on the surface to keep it simple: life is not rosy as it seems for everyone.
Well just want wish robin and keith a happy belated bday! =D
so where did i spend my 3 days?
Went to biopolis again for another genetics workshop. It's abt viral detection, for this case influenza.
Now before the normal stuff lets have some technical talk. ^^ The influenza virus is in short, our flu virus. If you have noticed, nobody seems to ever gain immunity from it or has ever developed a reliable vaccine for it. Ever wonder why?
Viruses such as chickenpox, measles and hepatitis B are all basically made of a protein capsid(de outer covering) and rna inside which carries genetic information. Their RNA is a long long strand that is linear. When the virus infects a host cell, this rna will be the one that instructs de host cell to stop producing proteins for itself but rather, make de viral parts and construct more viruses.
But in flu virus, the rna is not linear and it is in the form of 8 segments. Each segment control a particular part of the virus to be produced, such as the matrix protein in the capsid. But because the rna is arranged in 8 segments, there will be more chances for mutation when replicating in the cell. A minute change in any one of the segments will cause a change in the property of the virus, for example, its matrix protein changes. Then a new subtype or an entirely new type of flu virus is produced. This is why flu virus mutates so fast and why we cannot find a reliable vaccine for it.
So lets examine the classification of the flu virus. There are 3 kinds of flu influenza A, influenza B, influenza C
influenza A is the most diverse in all 3 families. It mutates very quickly, have high virulence(contagious) thus it has many variations and it infects both animal and humans. Alot of flu epidemics are all caused by influenza A: Spanish Flu Hongkong Flu Asian Flu Avian Flu
Influenza B almost only infects humans and the only other animal it infects is the seal. It mutates 3 times slower than type A and thus is less genetically diverse.
Influenza C is most least understood and unlike A or B, it has 7 rna segments instead of the usual 8. It infects humans and pigs and can cause severe illness. but it is very uncommon to find influenza C.
How do we name the flu virus types? we heard recently that the Avian flu was named H5N1. So how did they give it this boring name?
H is actually HA and is a glycoprotein on the surface of the capsid. It is called hemagglutinin. This helps the virus to get into the cell by binding onto the receptors of the cell. there are alot of types of HA glycoprotein, ranging from H1 to H16.
N is actually NA and is also a glycoprotein on the surface of the capsid. It is an enzyme call neuraminidase. It helps the virus to get out of its host cell after reproducing. There are 137 types and 9 subtypes of NA.
So if a flu virus has HA 5 and NA 1, then the virus will be called H5N1, which is your bird flu.
So now we know the structure of the flu virus, how do we see if a person has the flu?
1st we use a throat swab or phlegm sample and put it into a small eppendorf tube and let it incubate awhile. Then we add some enzymes into it to kill break down all the cells and virus in it into protein(from the cells and the virus coat), rna(from the viruses) and dna(from your cells). Then we add Dnase and proteases in it, filter, to get rid of the dna and the proteins and wad we left is the viral rna.
Then we do reverse transcription PCR(polymerase chain reaction) to amplify(make copies) the viral rna into large quantities of corresponding DNA for testing. Reverse transcription is the opposite of transcription. Transcription as we all know, is making RNA from DNA. Reverse Transcription is the making of DNA from RNA. Ingenious right? i know. =P
Why do we need DNA and not RNA in our testing? firstly, RNA is less chemically stable and will break down easily(as it is only one strand, the other side left open). Secondly, when we are to detect the virus later, we are going to use a detector called ethidium bromide that will only bind to DNA and not RNA. Thus, if RNA were to be used for testing, no results will be shown. Thirdly, to amplify the DNa we would use DNA polymerase, to amplify RNA we would use RNA polymerase. The difference is that RNA polymerase is less accurate in making copies then DNA polymerase, and will make alot of mistakes, causing inaccurate results.
so wad is PCR anw? PCR is polymerase chain reaction. It uses DNA polymerase, which replicates DNA, to create many more strands of a particular segment of DNA you want to amplify. In the case of reverse transcription PCR, it is amplifying DNA from RNA. There are a few components in PCR. Buffer Enzyme mix DNA/RNA template Primer mix DNA nucleotides
The buffer is actually basically water with magnesium ions in it. DNA polymerase is an enzyme that has an active site that is activated by the presence of magnesium ions. The higher the concentration of Mg ions, the faster the reaction, but the less accurate the product DNAs would be.
Enzyme mix is your enzymes required in the reaction. In this reaction the enzymes are Reverse Transcriptase and DNA polymerase(TAQ polymerase to be specific). Why Reverse Transcriptase? To perform reverse transcription you have to have reverse transcriptase, and then from the DNA produced you amplify it with the TAQ pplymerase. An important thing to know is the optimum temperature of which the enzymes work. Reverse Transcriptase works best in around 50 degrees celsius. TAQ polymerase on the other hands, is taken from a hot springs bacteria called thermophillus aquaticus. Thus it would only start working in high temperatures such as 95 degrees celsius, in which the RNA polymerase would denature.
The Primer mix we can say is one of the most important things in the whole concoction. Earlier said, we were going to amplify a certain sequence, only around 195 nucleotides, and not the whole strand of Viral RNA. To make copies of this particular part of the nucleic acid, we would not use enzymes and cut it or wad, as it would be impractical since enzymes are specific in where they would cut and in every PCR we may want a different section amplified. This is where the primers come in. The primers are single stranded DNA that are around 3 to 5 nucleotides in length. It nucleotide sequence is corresponding to sequence at the sides of the section you want to amplify. for example the DNA is ATGCG........GTACG, where the .... part is where you want to amplify. The primer would be TACGC and CATGC, where TACGC is the start primer and CATGC is the end primer. Notice that the start primer corresponds to the start of the sequence and the end primer corresponds to the end of the sequence.(note: A binds to T and vice versa, and C binds to G and vice versa too) The start and end primer would then bind to the part of the DNA or RNA to be amplified and serves as markers for the polymerases to start and end copying.
The DNA/RNA template is the RNA or DNA we want to amplify. For our case, the viral RNA we obtained by the throat swab. The more template the faster the reaction would be at the beginning.
The DNA nucleotides, or DNTPs for short, are the basic building blocks for the DNA(the bases and the phosphate backbone). The polymerases need these to construct new DNA, linking and bonding the DNTPs together to form DNA as they progress in the reaction.
So we add optimal amounts of all these together, go through several cycles of heating and cooling which is regulated by a machine, and finally end up with the product, alot of the particular DNA we want!
There were 3 specific places of the RNA we wanted to amplify, one was the H5 gene, second was the matrix protein for flu A gene, and the last was the matrix protein for flu B gene.
So we take the DNA produced and conduct a test called gel electrophoresis. So there would be 4 results possible. first is positive for flu A H5 subtype, second is flu A, third is flu B and fourth is negative. This is because we test for flu matrix protein gene and the H5 gene. The H5 gene is only possessed by the flu A but not flu B, and not all flu A has the H5 protein gene(there may be H4, H1 etc...). So for flu B you would not see the flu A matrix gene nor the H5 gene but just the flu B matrix gene. And for negative, it would mean you have no flu, or have flu C, which is very rare.
Just like that we will determine whether we have the flu virus. Like all experiments, this one also has a chance of screwing up, especially the PCR part. So we cannot trust the results fully.
***End of Techno talk****
Whew i didn;t expect it to be so long! the technical part actually postponed this post for a day! I had to type it halfway and save it to drafts to continue the next day... hahas okok so now its the normal stuff le.
Well as seen from the technical stuff, the whole thing is a boring and arduous process. Well got someone say before, its not the show that we pay for, but the company. Yes, make alot of new frens and they were very nice ppl. Curiously, the last time i went, the frens made were from VJC and NUSHS, and now, they are from VS and NUSHS, we close indeed. The NUS ppl also know kylie, hahas =PP
Well my lab partner, shes from india! her name is Sneha =DD The first half of bench one and two, lesser than 3 them Saw this funny poster on the wall, appears to be koped from talking cock The supervising scientist, we disturb him take his pic... hahas and he will tell us to sew lab coat buttons if we break the rules. He do his worm experiment, no relation to us. Liquid nitro! Went out with the VS frens for lunch. The lecturer. some guy from japan. He talk english very slow and jerky, but better than last time de peng jirong. no china accent. hahas. He very weird. He come to the lab to talk to us, then i letting my frens hear a song in my phone then he suddenly disturb and ask, "Can you live without phone?" then talk alot of crap shit. hahas then he ask my partner why she come to imcb and wad she want to be in the future. Then he ask wad why she take bio and why she like bio, walau eh so extra. Saw something funny in the mrt.... wad is this man doing? secretly fcuking the guy beside ah? lol Me and Jin Meng Vera! =D The gels left from the electrophoresis. Our legacy we left at imcb.... the KCPSS, RGS, RI, MGS one all very nice hor! All have my participation one eh! KCP of course is i draw la, MGS also i help them draw, RI i give suggestions abt the color and RGS i help them ink and color! woohooo =PPP The sereneness of the last day.... =( Future Doctor eh.... =X
Hahas so after all that went back to school on friday afternoon to meet dex and gang. then went to football. After everybody else go home, dex and I went J8 to eat and drink. Then went home. Chem wokshop today was fun... blog abt it nextime... no time upload pics from phone =P Heheheheheheeeh
Everybody's been through alot, so stop being so selfish
stop looking down
brightness will break through, and SHINE @ 8:58 AM
Saturday, April 12, 2008
hahas just had founders day... great success to everyone in our choir! =D it was an angelic performance of the "peace of god" and a heart lifting singing of the school hymn! Congrats, we made a sucky song sound nice. =P
Went to school at 6pm ytd, and was late. So i didn't have dinner. Then we changed and makeup(not me =P) and settled down. Now after being charmaine's assistant, i think i know how to put make up le! so who wan put can come find me =X
So we gone through the pieces and as usual, there was a delay (mr low like tok too long, as always) so we did our other pieces. 9.30 we sang! =D Racheal coughed once on stage but luckily no one heard. Xueying played the school hymn too fast because of nervousness... and somebody burst into laughter just before the performance started on stage! the sound like farting liddat sia.. we laugh alot after that.
When the performance finished, we change back to normal and went down where there was free food! saw mr atherton but didn't tok to him. Jeremy is like his bodyguard sia... even have the wireless thing on his ear, lol. The food was nice (as i was hungry) and i met zhiwei and hannah! they probably came for the prize giving. Jeanette and the other graduated choir seniors came to see us too. And for some reason, amanda also came. Hannah! Amanda teo Jeanette! =D
kk now i need go do hw le! ahhhhh ><
stop looking down
brightness will break through, and SHINE @ 4:00 AM
hahas so heres the concert.... This concert was organised by toh ban sheng and is named "i believe", starring Dunman High choir, Catholic JC choir and St Anthony Cassonian sec choir. It bore this title because mr toh ban sheng supposedly said the words "i believe" to the dunman high choir that spurred them to the state they are today.
Well the choirs are good, so is the conductor, mr toh. But many ppl hate him. Firstly, lets talk abt his good points. He is very dedicated to teaching choirs and sacrificed a chance of a professional solo career just for it. He may very well be a tenor or countertenor soloist and earn big bucks, but he didn't and chose conducting instead. Scondly, he has a vast knowledge of vocal pedagogy and experience in teaching choirs, even much more than mdm yeong. Thirdly, he has an excellent voice(as heard from jeanette) able to sing bass(maybe?), baritone, tenor, alto, mezzo and soprano. This makes his range to be around 4 octaves. His sopranino range is unheard of but it may very well be there(if it is then his range would be 5 octaves). I for one have experienced the power of his voice when he came once to teach our choir last yr. His soprano voice, sung in falsetto, can reach up to a deafening fortissimo while reducing to a light pianissimo as well, truly breathtaking vocal and breath control.
The bad thing abt him is his abuse of his ego. He that time come and teach our choir once, and he tell the school that he don't want to teach. He say that if he was to teach our choir, he would have to start from scratch. True, i agree that we aren't good, but how can one speak so foully? All because of ego. He also is too dedicated to choir that he set very high expectations to those in it. He holds 3 practices a week ending very late each time. And if near a competition, even tell ppl to sacrifice their church time and duties to come to practice. This is atrocious as it actually infringes on the right of religion. Lastly, he does not correct ppl's mistakes while scolding them, but tell them to figure out themselves, as opposed to mdm yeong. Proud and arrogant, with such a voice, it's no wonder ppl call him gay.
His mug, taken from facebook
So the went to the mrt station around 6 to meet farah and charmaine. Tehy were taking the mrt from angmokio to bishan then to city hall. I was supposed to meet them in the last cabin. But tragedy happened, i boarded the wrong train and go the wrong direction! =(( so i later then chase up and met them in city hall. then we reached victoria concert hall just in time for the concert.
The concert opened with a song sung by the st anthony choir we were very familiar: The peace of god. We sung that for our founders day. It was truly an enlightening experience, hearing the same song our choir is singing being sung by another choir. It reminds us of how much we still have to improve. Every dynamic was carefully expressed, every harmonization was carefully balanced, it was an excellent performance.
After that came legions of songs sung by dunman and cJC. the first song that had a soloist was the "circle of life" from the lion king sung by dunman. Farah was mesmerized by the guy's "sexy" voice(tenor). Sigh... Girls... =P The dunman choir was easily the biggest choir i've ever seen, with around a hundred members. I heard from mr niam their history was that when mr toh just came, it was as exactly like our choir! small with only one guy. hahas. Then slowly they changed and soared, achieving gold in various events. Now they are a SATB choir(soprano, alto, tenor, bass). All because of mr toh's "I believe". Hope mr niam can do that to our choir too, after all, he is mr toh's disciple.
Then the grand finale, all the choirs combined to fill the whole stage. It was a spectacular sight. Then the song was nice too. it also included various solos of bass, tenor and soprano parts. Then the male soloists came out and danced with female soloists and everybody all like "awww..." lololol so funny. Charmaine was head over heels abt a guy soloist also from dunman, because he very shuai... lol, she said that last yr dunman choir came to our school, she didn't have the guts to talk to him. hahas jiayous to her la....
So after everything ended, and mr toh went to the back stage, the choir was stil standing on the stage. The audience have alot of the friends of the ppl in the choirs, so they cheered and ask them to sing one more time. lol they insisted so vigorously that mr toh actually came out again to conduct for one last song. hahas joke sia... i dunno he so kind de. But then some of the ppl are like, "sian... why one more song! go home already la..." hahas =P
So after the concert ended, we headed home, on the way back we took some pics. hahas
I sort of miss mdm yeong... =( lol
stop looking down
brightness will break through, and SHINE @ 2:36 AM
Friday, April 11, 2008
tuesday i, andrew, dexter and shengliang went to library to study after tchoukball. It was raining and the second half of the match was postponed. When we walked there, we decided to take some photos. lol had fun.
f4
We breakdance! =D hahas andrew looks like he not in this pic, cos his face look very like robin in this pic... He jump super high la... then he had a great fall, lol, like humpty dumpty! his waist got blue black le... and he keep toking abt all the way to the library.
In the library, everybody all very sian and restless. we had no mood to study so we decided to try out the punching machine in the arcade. We took some pics in the library lift though, then an uncle came in... lol
Little Bois! =D We luv the littel gurls... hahas
Andrew was kinda sad so he vented his feelings on the punching machine and the first punch was an inhuman 125kg! mine only 65kg =.= sad sia.... musxt go practice! heehees... Then after that punch andrew hurt his wrist. waist and wrist all hurt liao! how? hahas... then dex and andrew played a dancing thing like para para but is use feet one. very sian, lol. then i had to leave them cos i had to go to meet farah and charmaine for some choir concert in victoria concert hall...
well thats all and concert next time post!
stop looking down
brightness will break through, and SHINE @ 10:03 AM
Monday, April 7, 2008
Hahas went to during saturday
Had loads of fun... but went to dexter's church first with andrew and to take part in a mind and body race... got sudoku and hula hoops all these stuff...
Then we headed to watch everland... the show opened with an ordinary and peaceful beginning, but quickly exploded into a dramatic display of action, plot, and laughter. Mr humpty dumpty (which i felt sad abt his demise), Wendy the dreamer ( played by lovely blessing), clown (by matthew bellido) and doll(by melody) were the talk of the show. The cast were well synchronized with their characters and pulled off an amazing performance. Ever be Everland! =D
Bubbles of your dreams? (touch them and they pop, dreams do fail you know =() Furious discussion, (Baiyu, "I support your opinion!" *thumps chest*) Stealing the shooting star... The trio (Wendy, Doll and Clown) Trio again The trio again again (told you they were good =P) Princess possum and knight Ben! Adversary... The battle of the tongues! Clash! the battle of the swords! Princess into submission Married to itchy! (tarantula) Finale! Ending with a dazzling brilliance that shall last in our hearts and soul for a long time
The soundtrack for the show was shockingly well done, although there was only one song that i heard a tenor part, kudos to all of you... Especially miss debbie wong, who wrote and composed all three songs as well as the script. =D
Then we saw mr atherton, our grandaddy =P Laughed ourselves silly cos of this guard XD The class... Miss tay with class guys and her bf =D Mrs Gan, She's Cool! Our A star Mass Cheok! Class guys Hanging in there! =)
After that we went makan sutra to eat and slack near the sea... It was tranquil and serene, ppl were emoing and romancing... hahas Then ppl left one by one until only left me, dex, robin, andrew, dion and desmond. So after much discussion we decide to head back to bishan and ton. Thats when we met anabelle and jordan. Anabelle left us shortly though, leaving us guys... We disturbed her through the phone in seven eleven, making the indian cashier go red in the face. Then we went robin's house and play ps2. but i dunno how play so i play guitar lol... Then andrew, jordan and desmond left us for home so we headed to play lan.... WE played CS and dota, all of which i was very noob, but it was a fun experience. When we left the lan shop, our dark accustomed eyes were dazzled by the bright sunlight: it wad seven plus in the morning! =0 So we ate roti for breakfast and headed home. By that time, cos of fatigue, dion and robin were high, joking and cursing and laughing... Well insomnia's their Ecstasy.
Emoing on stairs Rocking the stairway Our shoes and our woes Rabbit in Robin's house! =D
Well so went we went home, there was no straight bus to my house and i was STUCK between the junction to my house and amk hub! =( all the buses were going to amk and the only way of goign home was to walk, or to take a cab. Based on the events last night, one might confidently predict that i was bankrupt! So i was stranded! Oh no. So i made my way home slowly and by the time i reached the 2 busstop from my original position, i was like a zombie. So i flagged a cab and a kind uncle offered to take me home! =D I should report this kind soul to the taxi company to give him a reward, but sadly i forgot his name! =( singapore may be a fine and cold country(not literally) on the outside, but actually it's quite warm inside. It was an interesting experience for me =))
Tired for the lack of sleep, i woke up late on monday and slowly made my way to school. I was just in time and was not late. But sadly they decided to close the door of the hall to stop those that are diddly dallying did not quickly assemble from going in. I was stuck outside with those ppl, with robin included. fucking new policy... =( so we got a scolding and then went in after the national anthem ended.
After school we had a chinese compo test and i took my time in finishing it, resulting in an unbalanced essay with too big of a head and to thin of a tail; the ending was sloppily done while the beginning was choked full of elaboration. =( dunno will i excel in it or not... wrote abt extinction of species, with all those save mother earth crap rolled in. hahas then after that went to library to study with sl, dex and darren. Did our hw furiously and dex managed to clear out all his hw! shiok for him la... hahas... then we went KFC and i stared at his chicken so nice to eat sia... ahahs... then we became spas and took vids... had a fun time! =D
Dexter's cup! =0
My spoon! =P
hoped you all enjoyed them =D
stop looking down
brightness will break through, and SHINE @ 8:35 AM
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during saturday
Jireh