2nd Day (5/1)
Apart from having to do the DNA quantification and PCR, we actually forwarded our plan for the 3rd day, which included gel electrophoresis. DNA quantification, I think, is used to test for the approximate quantity of DNA obtained in your sample. First, we have to clean the plate, which in this case, we use Tecan's NanoQuant Plate (link here, video here) , then test if the wells of the plate are clean by a process called blanking. If the wells are clean, we then put a drop of the DNA sample into the well and measure it, all with the help of the machine and computer.
After that, we used the DNA samples from the first day (which we also used in DNA quantification just now) to do the PCR (Polymerase Chain Reaction). On that day, we just wanted to test for the presence of SRY gene (found on the Y chromosome) and ARG E1 gene. After going through the procedure, we then put the new samples into the PCR machine and waited for approximately 3 hours.
After putting the samples into the PCR machine, we went to prepare the gel used in gel electrophoresis. It just took a few minutes though, so Pong and I had to spend the rest of our waiting time day dreaming. When the PCR machine was done, we took the samples out and add a drop of SybrGreen (just a bit toxic) instead of ethidium bromide (carcinogenic) into the samples. Later we run the gel, and interpreted the results using a UV transilluminator. Here's a link to run the gel: http://learn.genetics.utah.edu/content/labs/gel/
3rd Day (8/1)
Actually we are supposed to be under Mr. Chia that day, but as the plans have been forwarded, we are now under Ms. Loo's supervision. She is a PhD student and is currently doing a research on stroke. That day, we attended two talks from two companies, one of which was Metasystems (I've forgotten the second one). The guys from Metasystems introduced their company's latest technology and software, which produces better results in karyotyping, , mFISH, mBand, etc. After the talk, we have free lunch (Haha~). Then we follow Ms Loo to run the gel with her samples. At noon, we had another talk from the second company, which introduces the SpeedCycler²,a machine used in Rapid PCR. Then we again run the gel with Ms Loo.
4th Day (9/1)
There were less activities that day, just PCR and gel electrophoresis with more samples and shorter time.
5th Day (10/1)
We repeated the same thing that day, PCR and run gel, it's just that the samples and reagents were different. However, that day Ms Loo showed us how to do the touchdown PCR, which is used to find the optimal conditions to amplify that particular sample.
6th Day (11/1)
The most boring and wasted day for the first week of the VRP. =_=
Our supervisor have taken a leave for two days (11/1-12/1) and we are temporarily attached to the cytogenetic lab. Actually I can tell that the staffs at the lab aren't totally prepared for our unexpected attachment, because we only had a briefing which lasted for an hour from 2-3pm (We are only supposed to be under their supervision starting from February). Our remaining time were spent by reading books, reading the lab's preparation manual, day dreaming and chit-chatting with another post-SPM student from CH, who is under Neuroscience. Sigh...
7th Day (12/1)
Today is a bit busier. We attended a lecture on Cytogenetics by an Indian professor, which is quite interesting. Later we went to the cytogenetic lab again, had yet another explanation, helped Mr. Chia with his new sample with a new polymerase, then back to the lab again to see the slide preparation technique. It's not any normal slide, but one involved in making out the karyotype later.
So, that's all for this week. (Damn, I've been spending 2 hours on this! =_=)
***
Hmm... I still have something in my mind which is bothering me now, though most of it have been solved by Panda. However, I don't know what it is that's bothering me. No even a hint! (Ha...) Hey, you're not the only one who's blur here, ok? Wakakaka~ @@
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