#theactualinternship

[nehsheh]

This week...

I haven't written anything for quite a while - sorry Amma and Dadda!:)  In any case, here is how my week went:

Work - learned even more and more, which is exactly what I came here to do, so no regrets. 1. Ethanol is god Since my lab is situated basically in the land of oil refineries and chemical factories, no flames. No bunsen burners, no flame-sanitizing things. Everything is cleaned and sanitized with ETHANOL. EtOH, FOOL! I must admit though, this is a little weird to me; by California law, ethanol isn't valid as a sterilizing agent. You must sterilize with bleach, and EtOH can only be used for superficial cleaning.

2. Cells don't die -  if you put them in a glycerol suspension. Glycerol is a cryopreservative, which means it prevents water crystallization in cells when you freeze them. For all my cheimaphobiacs out there (people with a fear of cold), hypothermia is damaging because it causes the water in your cells to crystallize, expand, and pierce your cell membranes. If you keep thawing your hands as they freeze and refreeze, this is what kills your cells. I asked if glycerol can be used to preserve humans or other mammals in the same way (without killing them). No, it can't. There are techniques, but this is not one of them.

3. If you open any media outside of a biosafety cabinet (BSC), it is instantly desterilized. Don't do that. (This one is also weird to me because my last lab didn't even use these. Glad I was exposed to this device!)

4. Harvesting Cells! Man, cells are just like babies. You gotta feed them and keep them warm and coddle them and treat them gently. Then you centrifuge them at 4,000 rotations per minute, freeze them in liquid nitrogen, and stick them in the -80. (Not the babies. Monstrous.) When we put the cells in liquid nitrogen, we heard this epic fizzling/burning sound, and my supervisor was like, "The cells must be screaming inside." HAHAHAHAHAHA

And now for play:

Dammit, random rotation! I definitely had this oriented correctly before I uploaded it... 1. Marche The CUTEST Swiss restaurant ever! Located just outside the Somerset MRT. All the decoration has this Swiss Alps/Alpenlebe vibe, and basically all the food is fresh (straight from the marche), and you just point to things and people cook them for you. It's AMAZING and delicious! I had a Swiss rosti, which is shredded potatoes cooked in a shizzton of oil, sour cream, and onion. Fattening, but SO GOOD! My Singaporean friend Risha's birthday is next week, but she's flying out of Singapore this weekend, so this was my last chance to treat her. She picked this place and I must say, good choice!

2. Ladies' Night in Singapore Of all the benefits my gender has, this one is among the coolest. My friends and I went dancing, and we got into the SWANKIEST club in town, Avalon - an impressive building of glass with chandeliers, the coolest ambient lighting, and view of Marina Bay. INCREDIBLE. Oh, and I got in for FREE. THASS WASSUP! The DJ played some hip hop, which made me happy, but majority of it was pop. I can still dance to that, so okay cool fine. Then we lost one of our friends, looked for her for two hours, stressed A LOT. She turned up the next day at work, so it was all good. I WISH I HAD A PICTURE OF THIS PLACE BUT MY PHONE DIED JUST AS WE GOT THERE! ARGH! I'll snag some off a friend and put them in a later post. Wednesday iskind ofa random night for Ladies' Night, but everything is free for the ladies - it really adds up to a good business strategy for the clubs, and a good win for me. "Something nightlife-y" is off the list! 

Finna catch a late showing of Dark Knight Rises and then head to Malaysia tonight. Fingers crossed!

[nehsheh]

List, work, stress, laundy

Waiting on my ish to dry. Ah, midnight laundry - can't escape tradition. Definitely owe my friend for letting me borrow his laundry card. Anyway, few updates:

1. I learned how to restriction-digest things. It's a way of cutting DNA at very specific places called restriction sites; once you figure out certain parts of the DNA sequence, you can manipulate the it with restriction enzymes (so much restricting up in here!). You can use it to make a circular DNA plasmid vector into a linear one, etc.

2. Night Safari - went to it as a child, revisited as an adult (or a 20-year-old. Whatever category that falls into.) Got the park hopper pass, so I can visit the zoo and bird park at some point too. Definitely was interesting. Although some of the attractions were closed, like the lion feeding (not done on Sundays. Apparently, lions don't eat on Sundays.) and "Suspense Bridge" or something, we still managed to see tons of deer, mousedeer, hippos, elephants, civets, tarsiers, a centipede (single. One is creepy enough.), a BAT CAVE (THAT'S RIGHT!) where bats fly by your face and devour fruits right next to your head, a flying squirrel, pelicans, crocodiles, otters, a porcupine, baby river cats, and more noctural things. Got some of that "I'm in Jurassic Park!" feeling, and the fun is only amplified with a group of 12-14. And walked away with a [stuffed] giraffe! Still gotta name him/her/it, though.

3. Was supposed to go to Malaysia this weekend, but reservations fell through and had a weekend of fun in Singapore instead. Between the lunches, dinners, and shopping, I didn't feel the lack of Malaysia too much. Ice cream and froyo (four times in the last 3 days. I'm stepping up my sugar and dairy game.), Arab St, Holland Village (which was a slight disappointment only because we saw the restaurants after we had eaten at the food court - FAIL). On a sidenote, I love that my guy friends will tolerate me pausing our mall walk to go shoe shopping. And that they actually ask me about what I've bought and give me commentary about it. Like actually. It's refreshing (if you're reading this, keep up the good work!).

Tonight, my bffl, my Turkish friends, and I were going home from dinner, but we stopped in Clarke Quay to see this Turkish ice cream place (and enjoy the night view); the person serving us had all kinds of cool slight-of-hand tricks. These two gentlemen got us our ice cream too! Super sweet :)

4. Trying to organize a stay at Marina Bay Sands (the place with the sick infinity pool), and after debating about days and prices, finalizing a day to go, sending out a thousand confusing FB messages, realizing I am not old enough to book a room even though I promised I would - I am ridiculously stressed out. Super glad that there are people in our group (like Alex) who are willing to step up and organize too. Irritated at myself and extremely stressed, but happy for the initiative. As a wise man once told me, "Just stay happy."

[nehsheh]

Learn ALL the things!

That is probably the meme I overuse the most. Anyway, I've had a lot of protocol and theory crammed into my head in lab over the last few days. I am actually SUPER happy about it too, though I get a little confused at times.

1. Running Gels I've learned how to make and run a gel to test the size and quality of DNA fragments; I loved doing this because 1. I feel like a real scientist, and 2. I used to see the researchers at my old lab doing this all the time, and I now know what it actually means/how to interpret the results of a gel.

2. Growing Things I've also precultured cells, streaked at least 32 things onto various agar plates this, and induced cells with methanol (poision for us, fodder for E. coli. OM NOM NOM.)

3. DNA and all that good ish Oh one thing I definitely wanted to share - extracting DNA from cells is far too easy these days. Honestly, every single thing you need comes in a handy dandy kit. All you have to do is pipette and transfer according to simple, stepwise instructions (which is really only as difficult as baking cupcakes. a.k.a. pretty freaking easy). I wonder what the pioneers of DNA technology had to go through for us to have it this good. Who discovered the use of buffers "P1, P2 and N3" (the kit labels; the fact that I have no idea what these are but can use them anyway means this is far too easy), figured out exactly what supernatant to keep or discard, and realized what those pellets at the bottoms of eppendorf tubes actually are? Makes me think of the whole "playing God" idea. Biotechnology is often demonized in media/movies like Spiderman (yes, I saw it last weekend and I liked it except for that one little aspect...); the possibility of that much manipulative power is far away, but I somehow feel one step closer to that myself. Now I'm just getting cocky, but it's an interesting possibility...Jurassic Park being my favorite movie and all (there is a link between that movie and biotech, I promise).

                                 It's definitely exciting to me that I now know how to do all of these things. Given a plasmid vector, I could inject and grow a cell with specific genes that do what I want. My lab group is running several experiments concurrently, which makes it a little difficult for me to figure out which protocol falls at which step in the experiment, but the more I do this, the more it will makes sense to me. Looking forward to it.

[nehsheh]

EFB and Gels

EFB = empty fruit bunches. That is exactly what it sounds like. They're abundant in Singapore, and make good substrate for some labs' projects. Picture, courtesy of Google search and the lucky first hit, etwau.com:

We got these in shredded form and spent some time grinding them down further; props to my colleague/co-intern for getting through them so efficiently.

And I got to watch how an agarose gel is made and used today. You make an agar solution, mix some toxic dye into it, give it a specific shape (combs/plastic squares with teeth create wells in the gel slab), let it set, then fill it with DNA fragments and some test controls, and run some electricity through it (Scranton! WHAT? THE ELECTRIC CITY). At some point, the DNA fragments separate by size and you can cut out what you need or assess your fragment quality. Do look forward to using this technique and have definitely stopped wondering why I kept finding comb-shaped gels in the sink at my last lab. :)

[nehsheh]

Recombinant DNA at last

On Friday, learned how to make cells with recombinant DNA! Given that the recombinant plasmid vector (I'm going to assume that's an actual term and not something I made up just now) was commercially available, i.e. we ordered that part from a company, I learned how to open up cells to insert foreign DNA into them, regenerate them, and culture them onto LB + antibiotic plates. I learned what electrocompetent cells are, how electrophoresis works, and how to spread cells onto plates properly. Also, I got to make the plates personally - my first love, lab-wise. I now know exactly what liquid LB, hot water baths, and shakers are for ("for what LB and hot water baths are," since prepositions aren't allowed to be at the ends of sentences); it feels good to have known what these were before, but I feel like even more of a scientist now that I know how to use them.

Recombinant DNA techniques are something I came here to acquire, and I learned part of it. Feels pretty good. Now I just wanna practice my technique and keep doing this; fairly certain I will have much opportunity to do so in the next several weeks, since this is basically my entire project.

:D

[nehsheh]

HEPES buffer!

Apparently, when water is purified, the metal cations used in the process make the water acidic (pH 4, ma nerds); we needed to use it to make antibiotic aliquots today, so the thing had to be neutral. Two frustrating hours later, my colleague and I go to lunch satisfied with our pH 6.97 water. Then, we find out that the aliquot can be make in HEPES buffer, which is super easier to pH - pleasantly surprised. :D I remember always seeing a box of HEPES in the chemicals room at my old lab, and I always wondered what it was. And now I know.

[nehsheh]

First actual day! And Happy 4th of July!

Got to actually do some lab work today! My supervisor is ridiculously busy, so I haven't been able to meet him yet, but I have a postdoc advising and teaching me. Today I learned how to do an assay - how to test for enzyme activity via absorption; don't know what it's called exactly, but I learned the protocol today. It was legit. My postdoc paused periodically to explain each step of the process, and a super chill PhD student gave us a lot of theory and good company in the lab; I definitely appreciate being given this knowledge, because this is what a real scientist/engineer uses to troubleshoot; one day when I'm doing an assay on my own, I'll be able to fix whatever goes wrong.

One tiny feeling of accomplishment I had today was that I actually knew what most of the things in the lab were/did; being lab assistant for a yeast lab definitely pays off when you're on your own yeast-related project. "Oh I know what an autoclave does! I know what a YPD + Zeocin plate is! I know how to use a micropipette!" Sounded good in my head, anyway. I know it's not much, but despite my frustrating mess-ups today, I felt good.

Also, chillaxed for the Fourth of July today. Happy Independence Day, America! Did the "typical American" things today - eat at Pizza Hut, eat at McDonalds. Pizza Hut is a sit-down restaurant here with gourmet food on the menu and star anise on the menu cover. We don't even have those in America...

Starting a whole new portion of the project tomorrow, so gotta prep myself for that. Starting with some sleep. Lehgo!

[nehsheh]

First day of work!

Finally stepped into the legendary A*STAR ICES (Institute of Chemical and Engineering Sciences) today. Missed my first bus and accidentally spilled all the private contents of my purse onto a service counter today, but I've learned to accept that I have awkward first days. Filled out some HR paperwork, met the PhD student and other lab members, learned about the project, and toured what is perhaps the most well-stocked lab ever. Definitely impressed, and can't wait to start! Gonna get my lignocellose pretreatment on!