air-cond down

Came in to work this morning with lots of plans for the day. Then remembered that I have to go update my paper work for summer payroll. Went to the Office of International Students and Scholars and was told that I need both my passport and I-20. So I came back to the lab, put some stuff into the autoclave and went to OISS. By then, my nose was already leaky and that kinda threw me off hook for the rest of the day.

Went back to Biomedical and Physical Sciences Building (BPS), where my lab is at, and was told that there will not be air-conditioning today due to some maintenance. It was so hot and the air was stagnant as all the doors are air-sealed to maintain separate air-flow usually but it's not very helpful for situation like this. So we had to switch off all the lights and unused incubators. Kwi and Steph tried to use the PCR machine and it gave some heat block error, which is probably due to the heat in the building. The temperature is about 31oC, which is a less than regular M'sian weather but I just could not take it today because I'm unprepared and I am still having a sore throat and runny nose. But oh well, I should expect it since I'm usually like that in M'sia but it's so much harder to work when I'm feeling crappy. It does not help that I still need to confirm the plasmid before I go back.

By mid-afternoon, we all agreed to a cool break so we went for a long lunch at Panchero's by Grand River Ave. It wasn't very cooling in there but it was way cooler than in the building. When we got back to the lab, it was even hotter because the sun is already up high and it's shining straight through our lab window and I know plants benefit from it but I surely do not like the greenhouse effect. At 6.30pm, I got 12 blue transformants shaking and incubating at 37oC and the 30 ml culture for fluorescence protein purification was still not growing yet. So I left for the library hoping to get some casual reading done but it was closed. I ended up going to MSU Union and sat by the window while reading my novel. It was very enjoyable and relaxing and cooling!!!

Came back to lab and spent the entire night organizing stuff and trying to pack my stuff in my mind. I think I just have a little too much stuff to carry back..unfortunately most of the stuff are not mine. I definitely gotta learn how to say NO to my brother when it comes to asking me to do stuff or buy stuff for him. But oh well..it's too late now.

~hot~

awful flight

We got to Atlanta International airport at around 12noon to find out that we could not check in because the self-service kiosk could not find out flight number. The reservation ticket we had says that our flight is a Northwest flight that is operated by Atlantic Southeast Airline but, we were told to go to a Delta counter. Lined up for about 10 minutes while bitching about how the airlines are screwing us up when a Delta airline assistant came by us and told us to go to the Business Class counter. Went there and we were told that the flight is there and he was surprised that we could not find it on the electronic kiosk. So he just gave us our boarding pass to Minneapolis, MN and told us that we just have to get to our gate at Minneapolis and we'll get the boarding pass from there.

Passed through the security in a breeze and had a quick lunch at Wendy's. Unfortunately, our flight was delayed due to tyre problems. About 45 minutes later, we were boarded onto the plane and it was moving towards the runway when we were told that we have to turn back due to air traffic. But 1o minutes later the flight captain announced that we'll be departing forMinneapolis. We didn't thought that we would make it for our next flight from Minneapolis to Lansing as we arrived at 4.15pm for the 4.45pm flight. We walked from Concourse E to Concourse B, which didn't look very far but it felt like the longest airport walkway ever. It's moments like this when I'm glad to have technology such as the speed walking thing (what do we call that anyway?). When we got to the gate, the flight attendant was kinda mean to us for not getting our boarding pass. We were all cranky and tired and that's what we get when we reached there. He didn't even want to look up for our names to check if we are on that flight on not. He just told us to wait until he finish boarding this plane so that he can check on the next flights. It was only until everyone has boarded and he realized that there are 5 empty seats that he started asking our names one by one.

I was so glad when we took off 10 minutes later than the supposedly departure time. I was overjoyed to go home. Unfortunately, when we got to Lansing Capital City Airport at 7.15pm as scheduled, I saw a rainbow when the captain announced that we are landing. BUT, before we got to land, the captain announced that we have to wait above the clouds because there was a thunderstorm. There were also lightning so we hard to circle above the clouds until it ceased. But after about 45 minutes of circling, the plane was running out of fuel so we had to detour to Kalamazoo. I thought that would not be so bad either since I can just rent a car and drive back and it would probably be about the same time as the plane is gonna take. But they wouldn't let us down due to security issue. So we just sat and bitch to each other in the plane. We were all hungry and even more upset by then. The air stewardess wasn't very helpful either because he just sat there. He could at least entertain us or something, which was what happened in Steph's plane when she was stucked in Detroit during the thunderstorm.

After another hour of waiting, we finally took of to Lansing. That was a very scary ride as we can see lightning flashing. They were not close by but it sure was scary when we're flying so high up. It didn't help that my stomach was feeling queasy too. When we landed at 9.45pm, it felt like that's the best thing that have happened to me for the entire day. It sure is not one of my best days. I hope I won't be having any problems on my way home to M'sia or when coming back here. I would be even more upset as I would be by myself instead of with a group of friends.

~tired~

stuffed with science

On Tuesday, which was the 2nd day of the conference, Steph and me did not have anything planned in the afternoon so we went to the World of Coca Cola, which was kinda fun but I thought it was too expensive. We had to pay USD9 to get in and it only took us about 30 minutes to finish the whole place. It is simply a display of all the advertisement for Coke and the evolution of it's bottle and cans. There is also a kiosk of free drinks produced by Coca Cola from different places of the world. Since I'm not a carbonated drink fan, I lost out on that as it is unlimited drinks. However, I got a sip of the Fanta Orange, which I used to like when I was a little girl before my 'gee ku' added 'Or Kau' (a type of brandy I think) to my Fanta Orange. I also tried some lychee soda from Thailand but it doesn't taste anything like lychee at all.

Since Underground Atlanta was just right opposite of WofC, we decided to roam around there. Like other parts of downtown Atlanta, it is full of people loitering around the area. However, there are lots of nightclubs that looked really fun but unfortunately, we wanted to go to the Division N mixer at the CNN center. Division N is the division for microbial ecology. Unfortunately, we didn't know anyone there except for my undergrad mentor. So we just ate, hang around for a while and left. The food wasn't that bad either - bbq pork ribs and chicken fingers. We then went to crash the Marine Biological Laboratory mixer. They had tons of drink tickets left over but as usual, I only got water as I don't drink beer or carbonated drinks..yeah, I can get a little too picky with my drinks sometimes. Stayed there and chat until about 11.30pm, went back to the hotel and drop dead.

On Wednesday, after the morning session, we were all crying for a break because most of the seminars by that time, are more or less the same. It's kinda scary to feel that way because we've always thought that we've moved forward since the last decade. Unfortunately, it is still clear through these seminars that we've only managed to isolate less than 90% of the microbial diversity that is out there. I was intrigued by the research on coral reefs. I didn't expect to such diverse community at an environment so harsh as this but surprisingly, it is. However, they have no idea how the community affects the environment or vise versa. We still have a lot more work that needs to be done. Advancing in genomics does not seem to help as much as we thought it should. We've come to depend too much on it that we stop looking at the cells themselves. We have ignored life itself - we depends on DNA so much that we got ourselves into a trap where we no longer knows how to draw the line between species. We have to start living the moment.

Wednesday afternoon is the MSU mixer. It was probably the most fun night in terms of being social. I didn't recognize a lot of people except for the Young lab, some from the Whittam lab and of course, the faculty members. There were lots of alumnis there too. Unfortunately, they only serve finger food but one of them is the Thai style chicken satay (which cocunut sauce and peanuts instead of our regular spicy satay sauce). There are also all kinds of weird cheese, some even tasted spicy. The Young lab left early for dinner but since they wanted to go hang out after that, I did not follow as I was feeling kinda crappy. So I just hang out there and it was actually worth it because I actually get to talk to this guy that I think is really cute.

We were in the same class - MMG 801 - during my first semester here at MSU. I never thought that he would recognize me because he hardly looked my way and we never talked at all during class. After the last day of class, I never see him again and I never did thought about him since I didn't know anything about him. During the mixer, we exchanged eye contact a few times and I was kinda glad that he recognized me since people hardly remembers me. While I was waiting for Steph to finish talking to Dr. Esselman, I was standing by myself and he came towards me. We had a good chat and I sure find out lots of stuff about him. So that was the highlight of my day.

~giddy~

Day 1 Atlanta

Yesterday was the first day of the conference and my itinerary was full. It started out with a symposium titled “To Grow or Not Grow: Tactics and Strategies of Uncultivated Microorganisms”. I enjoyed the first talk on high throughput cultivation and Dr. Schmidt’s talk on trade offs between power and efficiency. The talk on persister cells is also interesting. The persister cells are a variation of a wild-type that was treated with an antibiotic that it is susceptible to. It is not resistant to the antibiotic as when it is treated with antibiotic, the numbers will decrease to leave another batch of persister cells. It is a phenotypic variation of the same strain of bacteria. The two remaining talks on resuscitation of dormant cells and the great plate count anomaly was not very convincing to me. If the presence of cultivable variant is required for the resuscitation of the noncultivable variant, then why isn’t most of the late colonies similar to the early colonies? Why are they of different species or even genus? I still think there are more evidence required before the idea can be widely accepted.

Went to the CNN Center for lunch – had Black Bean burrito which was really good. The weather was really hot and humid, almost like Malaysian weather but not as burning. We then went to explore the Student Lounge and found out that they serve free lunch there. But since we already had lunch, we just got some desserts, which were really good – it was a rich chocolate brownie! It was still early after we finished so we went down to look at more posters and later went for the afternoon seminar session. It was about Marine Microbial Diversity, which I thought was very interesting. In fact, the seminar and the poster session for that afternoon had inspired me to start considering looking at aquatic environment. Furthermore, it might be more applicable in the future since Malaysia is surrounded by aquatic environment.

The afternoon session finished around 5pm and we went back to Steph’s hotel – Super 8 motel. We freshen up and walked around downtown Atlanta. We intended to explore the merchandise mart which we saw on the map but unfortunately, it seems like the building is some office building with displays of all the merchandise company they have around here. So we ended up just walking aimlessly around downtown and when we got to some area with lots of people loitering around (kinda like Petaling Street, Kota Raya area), we decided to turn around and go get some dinner because it was getting dark and we didn’t feel very safe there.

Choosing a place for dinner wasn’t that hard because most of the places closed at 6pm. We didn’t want to go to Quiznos, which seems to be at every corner here in Atlanta, McDonalds or Hooters, which would be very inappropriate because it was just me and Steph. So we ended up going back to Hard Rock Café. I had a Caesar salad, which was really good and wasn’t too filling.

So that was my busy Monday. Full of science but I had fun too.

~great time~

weekend in Atlanta, GA

It’s soooooo great that the Fairfield Inn of Marriott has free internet access. It’s like the greatest thing man have ever invented. It’s not the daily e-mail checks made me so glad to have internet access but finding directions to get around in Atlanta. I was so busy getting the YLC construct to be cloned into the broad host range plasmid pMMB67EH that I didn’t think about printing out maps and looking for directions to the places I want to visit. At the same time, it gives me a chance to catch up with stuff online that I’ve been missing last week while I was so busy.

Got here on Saturday around 4pm.We didn’t have any transportation to get around and cabs are too costly. So we just explored Sheraton hotel, which was where we bunk over with Jason and Alicia on Saturday night. Then we walked around downtown Atlanta to get ourselves oriented. The Olympic Centennial Park is really beautiful. It has a water fountain that shoots out from the ground and they allow kids to play with it. I thought this is interesting because I’ve seen similar kind of fountain in Beijing, China and they wouldn’t allow anyone to go near it. The fountain becomes a musical fountain at night, similar to the one we have at Mines Wonderland.

Passing the Centennial Park, we walk by CNN Center and Phillips Arena towards the Georgia World Congress Center, which is where the ASM 105^th Meeting is held. We then went back to the hotel and chill by the pool before we went out for dinner at Hard Rock Café, Atlanta. Since I haven’t eaten at any Hard Rock Café before, I think it’s a really great place. The food is not too expensive either although I wish there was more healthy choices. The best thing there is the music. They played all the old rock songs like Bob Marley, Van Morrison and Led Zeppelin.

We were sitting close to a door that leads to some audio control room or something like that. While we were eating, this lady came by and requested that we move aside as she has to get something. Then another guy came along and asked the same thing. The manager then came to our table, apologize and offer to give us two free desserts, which I think was really cool because this would not happen in Malaysia at all.

On the way back from HRC, we drop by the liquor store to get some booze – got some Bacardi and Vodka along with some fruit juice. Went back to the hotel and watched Harry Porter and the Prisoner of Azkaban. After the show, we made some drinks after and went to midtown Atlanta around 11pm. We didn’t know which club to go but saw some people lining up outside of Eleven50 and we asked around. We were told that we should definitely check out this club so that was where we went. It is the first club in the States which plays techno that I’ve been to. Had two drinks – margarita and cranberry Vodka but had lots of fun. It’s been a really long time since I get to dance this crazy. But it didn’t feel that crazy because I was still thinking about the conference and feeling guilty about having too much fun when I have so much stuff to do.

Woke up around 10am this morning and went for a swim. Chill by the pool after that although I didn’t feel like it because the sun was coming up and I am definitely not looking forward to being dark since I’m going home soon. But I didn’t really know what else to do so I just hang out by the pool side with Heather, Alicia and Kacey but I was at the shaded area. Around noon, helped Kacey to check in at Westin Hotel, which is where she’s staying. She got some undergraduate award that allows her to stay at expensive hotels which are reserved the ASM. Went back to the hotel after that to take a shower and just hang around in the room until 3pm to check-in to Fairfield Inn. Once we checked in, we walked to the GWCC for registration. Before we registered, we walked over to the CNN Center to grab some lunch as we had not eaten anything the whole day. Walked back to GWCC to register and went for the opening talk on symbiosis.

After the talk, there was a reception where there are tons of food to eat and some bands and gymnasts as entertainers. The food was okay but as usual, it’s not very healthy choices. Stayed until about 9.30pm and came back to the hotel. So that was my busy busy weekend..it was fun but part of me wish that I’m back at Lansing working on my construct.

~fun~

going off..

*phew* What a hectic week. Lots of stuff has happened since I last posted an entry..in fact, so much that I don't have the time to post any entries or buzz around at other blogs I enjoy reading. Still busy right now..packing for Atlanta. I'm finally going!! Have been waiting for this trip for quite sometime and was actually not looking forward earlier today as I have so much more that needs to be done before I go back to Malaysia, especially since good results are coming, I am tempted to keep the tempo going. But who knows, the conference might actually be a good outcome if I can find someone to talk to about quantifying the fluorescence proteins I am working with.

Will update more when I'm back from Atlanta..as for now, I'm back to packing and planning the itinery for the next couple of days.

~anxious~

nothing's done

It's been busy these few days but I felt like I've done nothing. Lab notebook is now a few pages thicker than Friday but there are no long paragraphs written in green ink, which is the color I use to write discussions and progress. There are tons of paragraphs written in blue ink, which is the color I use to write why the result came out that way and what could have gone wrong. There are just too many ways things could have gone wrong. So what's wrong?

When the PCR worked, I was overjoyed. I tested 20 µl reactions for several temperatures and I decided that it worked best at 65^oC. So I decided to increase the reaction volume to get more products -- 100 µl. However, I did not multiply the DNA polymerase by 5 because I felt that it was too much polymerase and I can’t afford to use that much since 20 µl of polymerase costs USD22. For each reaction, I would require 1.5 µl polymerase if I were to multiply by 5. That means one tube of polymerase would not even allow me to run 20 reactions…too freaking expensive. No wonder Invitrogen can still afford to hire during the recession period in US. Unfortunately, the lesser amount of polymerase I used gave me a different kind of product, one which I don’t need at all. So now I have to change the conditions to get the product I want and this is taking a whole lot more time. There’s only 1 week and 4 days left to ASM. The fear of not being able to finish is eating me up.

While I'm running all these manipulations, I'm not progressing because I have yet to get my product. Sad. It really does feel like nothing has been done. Need to hurry..

~worried~

simple and fun

On Friday, my plan for this weekend was to watch DVDs, sleep in and wake up without an alarm clock, read novels and read some articles on microbial diversity. However, I could not sleep past 9am even though it was dead quiet when I woke up except for the sound of the cars passing by on the highway once in a while. But that wasn’t the source of my waking up. It was more due to the habit of waking up at 6am every morning and all I need was a couple of hours extra. But I got the pleasure of just lazing around the house, watching Hero and Jersey Girl.

Unfortunately, it is impossible to leave lab work even for a day so I went to lab in the late afternoon to run a PCR. Yeap, the PCR is working well but I still can’t get the product I want to amplify in large amount. It seems like the primers have another annealing side that keeps giving me an unwanted product that is only about 800bp. But I am glad that I have products to play around with instead of just empty reaction tubes with just buffer and water.

Around 6pm, CY called and said that they are having a barbeque at Hawk Island County Park. What else would be more fun on a Sat. evening than spending time with friends by the lake, eating BBQ chicken drumstick? So I agreed to it and he came to pick me up. I am glad that I went because we had so much fun and it’s been such a long time since I laughed so hard and it was genuine. I didn’t have to pretend to be a different person. The gathering was nice too because there was no gossips, which would have occurred if I had spent time with M’sians from my undergrad peers. I am not trying to offend anyone but everytime there is a gathering, there’s always stories going around about who’s doing what and what is happening to who. Here, we don’t know each other very well except that we’re all M’sians and we appreciate companionship of our own kind once in a while. So it was just enjoying the moment and nothing more.

After the BBQ, we had a few games of bowling at Okemos and pool at Owen Hall. I came home late and did not get to read much stuff but it was fun. This is actually the first time I went out with them as friends and not because it is organized as an M’sian Student Organization activity. I would definitely do it again.

line

I've been chatting to BK quite a bit these days. He broke up with his gal and he's heartbroken about it although he's handling it well. We don't usually chat much about he and his gal but more on general stuff..what's going on..teasing each other..just casual chat. But sometimes the topic would go to his relationship with his gal and how he's trying not to think about it. I listen and soothe him with encouraging words. Am I affected? Well, at least I'm not longer crying over it and losing my mind but it still hurts. It hurts because he's telling me how much he's affected by his break up and yet, when we broke up, it was nothing to him..at least that's the impression I got. So should I listen and bare with the hurt or should I just ask him to shut up? Where do I draw the line? As a friend, I don't want to see him get hurt and I don't mind being there for him..but hurting myself? Is that worthit? I guess I kinda know the answer to all this question in my heart but I just had to let it out. I hope he'll have an easier time letting go of her..surprisingly, not because he would be available again but because I don't want to see him get hurt.

Thankfully the experiment worked today. At least I have something to rejoice about instead of being miserable over my uncontrolled feelings.

water!!!

For the past two weeks, I've been working on amplifying a gene encoding three proteins fused together - yellow fluorescent protein:beta-galactosidase:cyan fluorescent protein - by PCR. Unfortunately, all I've been able to get were empty gels when I ran the PCR product on the gel. I've manipulated several conditions and components of the reaction and still came up with nothing.

Yesterday, Dr. R suggested that I try using the DNA Grade water from Fisher for the reaction or get a new batch of dNTP mix from Invitrogen. I did both but to amplify the 16S rDNA gene from Verrucomicrobium, which is a positive control and guess what..the reaction with the DNA Grade water worked!!!!

Just before I left work today, I started the reaction to amplify the yfp:lac:cyp gene, which was my initial objective. Will only get to see if it works tomorrow but I have a good feeling that it'll work. Can you believe that something as simple as water can screw up experiments for two weeks??? I autoclaved my water for 30 minutes and aliquoted it out into sterilized Eppendorf tubes and never did open it again until I need to use it. So the only possibility that's screwing up the PCR is probably some mineral residues in the bottle during washing or there is an inhibitor in the bottle and it was not washed thoroughly.

I can't wait till tomorrow to check it but I'm soooooo glad I find out what's wrong with the reaction for positive control.

~ecstatic~

Blood Red Tulips

Blood Red Tulips, originally uploaded by lilsQuirr3l.

First time I've seen this color of tulips, my favorite color too. This was at the Tulip Festival at Holland, MI. It's about an hour west of Lansing and the Tulip Festival is to celebrate the time when all the tulips first start to bloom. This was at a small garden and there's more pics taken from the Tulip Farm. Wanted to buy some but completely forgotten about it when we were about to leave. It is such a beautiful flowers and the variety of colors, I never thought it's possible to have so many types of varieties for a particular flower species.

weekend

Went to Ohio this weekend with Nok. The main purpose was to attend Ben’s convocation and Ben had free meals at a really classy Japanese restaurant. She told us that we had to be in Cleveland by 6pm. We got there a little before 6pm and called her. She was on her way back from Kent. So we waited in the car for about an hour and she called back, saying that she had already moved the reservations for the restaurant from 7pm to 8pm. That made Nok a little angry because she doesn’t like to wait around for people even if it’s her sister. Furthermore, when Ben said the original time she booked was 7pm, it frustrated her even more because we were not informed of that either. At that time, Ben was already close to Cleveland but Nok was so angry that she didn’t want to attend the dinner anymore. So we went to an Asian store at downtown Cleveland, which was about 15 minutes away.

At 7.30pm, Ben called but she didn’t want to pick up. So I talked to her and told her that I’ll try my best to convince Nok to go to the dinner. I tried to talk to Nok and said that it’s Ben’s graduation that we’re celebrating so she should try to be nice and cheerful. Told her that she can get angry after the convocation. She decided to go but took her time shopping. We arrived at the restaurant late but thankfully, it was so full that the table that was reserved for us was still not cleared. We waited about 10 minutes before we could get our seat.

Oh boy, that sure was an awkward dinner we had. Nok was extremely quiet because she was still angry. Ben on the other hand, was trying to make peace with her sister and entertain her friend who flew to Ohio from Texas just to attend her graduation ceremony. But I finally managed to get Nok to participate in the conversation and she cooled off towards the end.

On Sunday, we didn’t get a chance to shop either because we passed by Legacy Village, which is a high class shopping boulevard. We saw Cheesecake Factory and California Pizza, which was very tempting. So we ended up spending most of the morning there, went for Ben’s graduation and came home. So much for planning…which is why I prefers spontaneity.

When we were alone on our back here, she said that I am very patient and does not have a short temper. What can I say? I grew up with a rascal that always wanted to win in every way. I could have complained about him but it would not work because he knows his way around my parents. I could have hit him back but it would not work either because he tends to be a bit dangerous at times. There were lots of times when I had to sacrifice days of studying or spending time on stuff I like to do just to make him happy. Everything was about him. So I learned that the best way to stay with him in peace is to suck it up. That’s how I became so patient and angry is a foreign word to me.

~satisfaction~

still stuck

I'm still stuck in the first step!!! There's two more weeks to ASM and I'm still struggling with the PCR. I definitely do not want to leave it hanging before going home to Penang. Will I make it in time for the wedding? I sure hope so because I don't want to leave the DNA naked at -20^oC. It’s going to deteriorate and by the time I come back, I might have to make new ones. That means all my work for one month would be gone just like that. I do need to complete the plasmid construct before I go back. Need more time!! Need to work harder!! *Red light flashing madly*

Now waiting for a PCR reaction with less amount of template and Taq polymerase. According to Kwi, I might have added too much polymerase since it also has 50% glycerol, which might affect the reaction. However, I am already using way less than 10% of the reaction volume. This is so frustrating. It’s not like I haven’t done PCR before. I’ve done it countless times and I’ve even troubleshoot it for different kinds of amplicons and templates when I was working with Sinorhizobium meliloti. Unfortunately, I have yet to work with such long amplicon and it seems like this is a lot harder to troubleshoot. Anyway, I am also running a positive control, in which I use a Verrucomicrobium genomic DNA as the template to amplify for the 16S rRNA gene. This should work since it works for everyone and I’ve used the same primers to amplify 16S rRNA gene of H. hepaticus when I was in the Young lab.

Gonna go eat my dinner for now and hopefully one hour later, I will get some products in the reaction tubes. Will have to wait till tomorrow to run the gel as the last bus is at 10pm and the gel is gonna take at least two hours.

ants

Ever since I had dinner, which is curry chicken again, I kept thinking about what late ah kong (my maternal grandfather) used to call me. He was the only one who called me that. No one calls me that anymore.

Somehow, the thoughts came to one sentence.

"U chi chiak ang hia ka chi liap bee"

(there is an ant who is biting a grain of rice)

It's part of a story ah kong used to tell while I was a little girl. Unfortunately, that's all I could remember and it keeps playing in my mind because I'm trying so hard to remember the story. It's so sad. I wish there's a way for me to freeze all the memories of me and ah kong spending time together. Maybe I can ask ah mah about the story.

~missing you~

stuck

Sad to say, the PCR did not work again. I have decreased my template to only 10 ng. All the annealing temperature I tried did not work either. Sigh..

I think I should try to cut the plasmid first and use the restricted plasmid as the template. Maybe it was just too complex to denature completely. Furthermore, my expected product is more than half the size of the plasmid.

oh well..will try again tomorrow with any other approach that I can think of. Time to go home to eat curry chicken!

~disappointed~

PCR

I'm back full time in Dr. Schmidt's lab. Last week was a short break for me since it was finals week and a few days of just looking through my previous results from the Schmidt's lab. Had a chance to go to Holland on Sunday with Nok, which I might blog about it later if I find the inspiration to. It was fun because we didn't plan it at all and I love spontaneity.

Ever since I came back to the lab on Monday, I've been doing nothing but running Polymerase Chain Reaction (PCR). I have to amplify a 4.5 kb long construct out from a 7 kb plasmid. I've tried a few times with different conditions and I haven't manage to see even a tiny bit of product yet.

1. Used regular Taq polymerase from Invitrogen.
2. Increased elongation time to 5 mins.
3. Used XL rTth DNA polymerase from Applied Biosystems.

Now running one with annealing temperature between 45oC to 64oC and I decreased the DNA template to only 10 ng. Hopefully at least one of these reactions will give me a product. Something for me to move on. This is only the first step in creating the broad host range construct and I'm already stuck. Really need to get this done before ASM since after ASM I would not have much time before I go home.

~frust~

coming to life

Flowers are blooming and the surroundings seem to come to life the moment the weather starts to warm up. It's no longer white and gray but full of colors. Cheerful colors from pure white to bright yellow to fiery red. That's the best thing about spring. The depression seems to just die away and be replaced by the need to be out there, enjoying the wind blowing against my face, to feel the petals of flowers dropping from the trees when the wind blows hard, to just listen to the songs of the birds and calling of the goose. There's a meaning to everything, unlike the cold, dull winter that sprung the loneliness often too many times. It's definitely a moment to be treasured. I couldn't stop taking pictures and everytime I press the shutter button, I wonder how can I capture the sounds of nature along with the beauty of it. Of course, it's impossible so I would just have to enjoy it right now. But I just can't help wishing that I could capture every detail of the moment for my parents to experience it.

~peaceful~

all done!

Finally handed in the final paper at 2.30pm today. Finished it around 10.30am but was not very satisfied with it. Unfortunately, I had too much of bile for the past few days that I can't seem to think anything else different from what I have written. So I took a break and resume editing at 1pm. E-mailed it to the three professors who will be grading it at 2pm and gave the hard copy at 2.30pm. Could I have done better? Possible but at this point, there's no point delaying it until the very last minute to hand in since I can't seem to make any major changes anymore. Nevertheless, I feel so relieved when I handed it in without having to rush.

Came home after that since I have no mood to start any experiments. The lab bench is still occupied by the previous student anyway. Since my desk was in such a chaos when I left it this morning, decided to give a good cleaning. Organized all the articles into EndNote and I realized that I have read 169 papers throughout the past 3 months. How did I do that? It's more than 1 paper a day and most of them were read critically enough to make comments and create hypothesis for follow-up experiments. I'm amazed at myself. It didn't feel like there was so much to read but now that it's all neatly piled up on my desk, it's definitely thicker than any text book I've ever read. But I'm glad I did because I've learned so much about microbial pathogenesis, from the mild Haemophilus ducreyi that causes chancroid to the deleterious Escherichia coli O157:H7, which is responsible for a deathly bloody diarrhea, and from oral pathogen Phosphyromonas gingivalis to gastrointestinal pathogen Salmonella enterica and even to sexually transmitted Neiserria gonorrhea. It is still overwhelming to be absorbing so much data within three months, especially on areas that I am not doing research on but it is better now than later.

As of now, I'm just enjoying the few precious hours of uninterrupted break. Time to start opening the covers of novels...

~relief~

soon..

It is a little less than 24 hours to the dateline of this final term paper - Utilizing Bile as an Environmental Signal and Bile Resistance Mechanism - is due. I am a little less than halfway through writing it and yet it seems so hard to start this next paragraph. I can imagine myself rejoicing tomorrow at this time, cooking up a healthy tasty meal that I haven't had in a while and enjoying a novel I've been itching to start along with the meal. But yet it feels like it is so faraway when this paper is still not completely written and edited. I've been reading about nothing but bile and its interaction with the bacteria for the past few days and why am I having such a hard time putting it all together in my own words? I just took a look at the last review paper I wrote, which was about 2 years ago - The Green Fluorescent Protein as a Reporter Gene - and I can't believe that I actually did so well in that considering that I'm still stuck with this paper. Will it be as good? I don't know but I sure need to finish it soon or else tomorrow will be a disastrous day.

~overwhelmed~

enough!

Got a paper due on Streptococcus agalactiae in newborn invasive disease tomorrow at 1pm but I'm sitting here in front of the PC reading people's blog. Why? Distraction is the answer. I started out surfing on Web of Science to look for articles that discuss about the evolution of the Type III antigen strain of S. agalactiae, which is commonly found associated with newborn invasive disease. Then I got to PubMed to extract these articles and click the print button to print the articles. While I was waiting for the article to be printed, my hands flew across the keyboard typing penangfaces.chanlilian.net. Why? My mind was hungry for something close to home. This frequently happens when I'm stuck with a thousand things on my to-do-list and yet still have to think about what to cook for lunch because I am against eating out all the time. Guess what I saw when I first get to the page? Cream crackers!!! The first thing that came to my mind is cream crackers top with bak hu (chicken floss). That's my grandma's favourite food but at this time, I'm craving for it. So what's for my lunch? Sambal hae be with saltines. That's the closest to home for now. By the way, proud to say that my sambal hae be is brought all the way from my ahmah's kitchen in Batu Feringghi. It was brought here in a huge empty Greenfield Organic bottle and now there's only a little left but it shall be savored to the very last speck.

As of now, I have enough of papers to deal with. I can't wait till the last paper is hand-in next week and then it'll be free of papers to write for the next few months!!

~tired~