Day 22: On Gel Chrom

It's May 5, Day 22 of my thirty-day series.


I'm in the lab right now and today's agenda: Swelling of Sephadex G-100 and column set-up. 


For those of you who have no idea what Gel Chrom (Gel Filtration Chromatography) is about but wanna know more about it, click here.


Actually, Sephadex G-100 takes about three days (at room temperature) to swell BUT this swelling time may be shortened using a boiling water bath. Bottom line: my three-day waiting time is reduced to 5 hours, yeah! And now, three of the five hours have passed. Two more hours to go.

Day 14: Swimming

The usual centrifugation, SDS-PAGE and Bradford assay... nothing new.


The highlight of my day would probably be the overnight-swimming with my family. A little confession coming up. I'm already 19 y.o. but I still don't know how to swim. But hey, it's ok. I know that I'm not the only one who doesn't :P

Day 13: On Dialysis

Today's agenda: Dialysis.

I've done this for the third time already and here are the steps that I follow in dialyzing my crude protein extract.

Step 1: Wash the dialyzing membrane with distilled water.

Step 2: Soak the membrane in boiling water for five minutes.

Step 3: Wash the membrane with distilled water again.

Step 4: Tie one end of the membrane with a cotton string.

Step 5: Fill the bag up with your sample.

Step 6: Tie the other end of the bag with a cotton string.

Step 7: Suspend the bag inside a container filled with the dialyzing buffer.

Step 8: Place the container inside a bucket packed with ice.

Step 9: Stir the dialyzing buffer using a magnetic rod and stirrer.

Step 10: Allow dialysis to proceed for several hours.

Step 11: Change the dialyzing buffer then resume dialysis. Change the buffer after every several hours.

Step 12: Allow dialysis to proceed until the required amount of time is achieved.

Day 12: Merlin

Yes. Merlin, the TV series, is what I am up to right now. And I'm lovin' it. 

_________________________________________________

Here's a little bonus for all you reading me (except for my thesismates). 

A FREE MEAL for the first one who correctly guesses the two things I did in the laboratory today, April 21, 2010. Of course, these "things" have something to do with my thesis. And YES, a free meal to the first one who guesses correctly. I promise. Comment your answer(s). Good luck! [Bonus valid only until 7 pm, April 22, 2010.]

Day 11: Sick

Day 11. 

Nothing exciting happened today. Bradford assay here, SDS-PAGE there. Bradford and SDS-PAGE over and over again. Nothing new, really. Sometimes when you do something over and over again, you just get sick of it. BUT I cannot afford to "get sick of it." No, not now.

Day Ten: Garfield

Day 10. One day wasted. If there's one word that would best describe myself today, it would be "unproductive."  If there were two words that would best describe myself today, they would be "very unproductive." You get the idea... I was just not in the mood to do anything today. Maybe because it's Monday. Monday is like my bum day, if there's such a thing. Every Monday I feel so tired and sleepy and lazy.


Call me Garfield 'coz I hate Mondays.


I need a heavy dose of discipline here!

Days Eight and Nine: Donuts

Day Eight post came out a little... uhm... late and I'm sorry for that. These past two days went by in a flash and I've been very busy and tired. 

Day 8: Dialyzed sample for 24 hours and prepared a total of 6 liters of 10 mM β-mercaptoethanol.

Day 9: SDS-PAGE and Bradford Assay.


And now, to sort of "reward" myself for this week's accomplishments, I bought myself my favorite bavarian- and strawberry-filled donuts! YEAH! Am planning to buy some DVD's later, too. Happy weekend, everyone!

---

What's with the short posts? Hahaha!

Day Six: Pain

Day Six. 

My whole body's in pain. Clearly, trekking and hiking... not for me! I just wanted to lie in bed the whole day but, of course, I couldn't. I had to go back to the lab and grind my 16 grams of dehulled mungbean seeds.

Before dehulling:

After dehulling:

Grinding mungbeans using a mortar and pestle, for me, is more difficult than dehulling using a scalpel. I spent about three hours grinding and I hated every minute of it. Good arm exercise though, try it!

Short post. Tired. Wanna go back to bed. Whole body still hurts from yesterday's adventure. Arms, back, legs, feet. Ouch. Can someone lend me a wheelchair?

Oh, and I so can't wait for this week's episodes of American Idol. Adam Lambert as mentor and he will also be performing on results night! Something to look forward to , eh? :D

Day Three: Yet Again

Day three, SDS-PAGE (yet again). Checked on my 2nd electrophoretogram (from Day Two). It's better than the last one (from Day One), no bowling alleys but still not good. :(

Anyway, today's SDS-PAGEing went quite well, probably the result of our two days of practice. I haven't seen the output yet (destaining still in progress) but I sure hope it's better than the last. We'll see on Monday. 


Am not labbing tomorrow. Holiday. Long weekend ahead but I will keep myself busy by dehulling mungbeans. Mmmm... fun! 


Short post much? Blame this major headache!

Day Two: The Bowling Alley Phenomenon

Of the 24 hours of Day Two, nine hours were spent in the lab. First thing I did was to check on the now-completely-destained electrophoretogram from our first SDS-PAGE attempt (see Day One). What it looked like? Hmmm, let's just say that we have discovered a new electrophoretogram phenomenon. The people in the lab called it The Bowling Alley Phenomenon. Here's a snippet of the actual electrophoretogram.

Bowling Alley Phenomenon, get it?


Anyway... Today's agenda: SDS-PAGE attempt #2 and Bradford assay attempt #... uhm... Bradford assay, period. Bradford assay, quite successful ---> proof that, indeed, practice makes perfect. Our assay was not yet perfect, though. But we're getting there. :) SDS-PAGE? Failure (yet again). But as I've said, practice makes perfect. We'll get there sometime. :)


Tonight I'm gonna resume my Heroes Season 4 marathon. Or maybe I'll just finish reading Percy Jackson: The Sea of Monsters. Whichever I choose to do, I'll do it while eating Bavarian-flavored donuts.


Tomorrow's agenda: SDS-PAGE attempt #3 and summer 2010 registration.

Day One: SDS-PAGEing

Day One.

Woke up to a HOT morning thinking, "Must catch the 10:30-am shuttle to IPB. Must catch the 10:30-am shuttle to IPB." As I was getting dressed, I imagined myself looking at an electrophoretogram with perfect bands and without a single tear. Hey, I can dream, can't I? I have never PAGEd anything before but I had this impression that PAGEing would be such a hassle - from preparing all the necessary solutions, to preparing the gels, and to very carefully removing the ultrathin (exaggerating, of course) finished product from the glass plates.


Anyway, it was... uhm... not as hard as I imagined (maybe because the necessary solutions have already been prepared) but I'm not saying it's easy. I know I'll master the art of SDS-PAGEing (uhm) someday (?)


So, today's five hours were spent in the lab. And the fruit of our labor? Well, let's just say it's not the perfect electrophoretogram I had imagined. There was a "minor punit" and I haven't seen the bands yet (destaining in progress) - that's something to look forward to tomorrow.


To celebrate Day one, I took some pictures of the place that will serve as my 2nd home for a month. The Institute of Plant Breeding (IPB). 

I could get used to the view. :D