Wednesday, July 8, 2009

Day Three : PCR and Gel Electrophoresis

Day by day, my internship is getting challenging. Well, i've been waiting for the rest of my life (*if i can be a little bit exaggerating it^^) to learn about PCR and Gel Electrophoresis.
At first, i was a little bit worried whether i can succeeded doing those two things or not. Well, because this is my first time. But, having Mia as my supervisor, everything seems to be easy^^. FYI, Mia can't speak English fluently. But she is always very helpful and explains things i don't understand slowly (*using gestures, pictures...everything^^). Besides that, she's always one step forward in preparing additional readings for me. Printing or copying some lab protocols and references.
Back to the PCR and Gel Electrophoresis. After saw Mia doing those two, i feel like i couldn't do it. Quite complicated, indeed. Need patience, of course. Took some times for me to finish the whole PCR protocols. Dealing with the micropipettes are definitely easier than yesterday's toothpicks...hehehe. But still, skillful hands are highly needed. Finally, after couple hours, i managed to finish all the sample preps for bacteria primer and archea primer. Whew^^! Took 3 hours to run the PCR. During that time, i went out for my second yeast genetics class.
During the summer course class, i fell asleep again. OMG...since when i became Sleepy Dwarf??? I guess i'm lack of iron. Because the night before, i had enough sleep, so i don't think the problem is lack of sleeping hours. *better consumes more high iron content vegetables, Bung!*
Btw, the topics in this second class were getting deep. Those were difficult, really. Started from the yeast nomenclature, yeast genes characteristics, also yeast and cancer. I know if i don't try to catch up the topics and gain my understandings, i'll left behind. So, GANBARIMASU^o^!!!

After finished the yeast genetics class, i rushed to the lab and continued my works. Now, time for Gel Electrophoresis. Well, because before i have to go to the class, Mia has already prepared the agarose gels. Things i have to do was putting the PCR products to the gel wells. Wohohoooo...i liked this part very much^^. Difficult, yet interesting. After that, the electrophoresis chamber has to be connected to power supply. Because DNA is a negative charge protein, so later, we will see it migrating through the gel to the positive charge side. After 40 minutes, the gels have to be put to the Transilluminator (ultraviolet lightbox) which is used to visualize ethidium bromide-stained DNA in gels. The Transluminator was connected to the computer. So, we can see the data using computer software. The result showed that my unknown microbe was apparently an archea, not a bacteria. Because the bacteria marker-contained gel, showed only the marker band, no sample bands at all. While for archea marker-contained gel, the marker and the sample bands are all clearly shown. Yeay^^...first attempt, succeeded. Alhamdulillah...

See here for the PCR principles and here for Gel electrophoresis procedures.

Now, i have to deal with one thing. I'm in love with PCR and Gel Electrophoresis. Hehehe... *addicted_mode_on*

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