Day 5 - What's wrong with PCR today?
Day 5 - What's wrong with PCR today?
The first thing that we did in the morning when we arrived in the lab was to run a gel for our PCR (genotyping) result. We did not get the band that we wanted. Instead we got a single band for each lanes on uniform size, the size of the primer dimer, which means that our PCR result is not successful. Thus, Hong Ni asked us to redo our PCR.
We repeated every step, but this time we made our own PCR master mix, and run a gel again. At the end of the experiment, I still got primer dimer but Shen Qian got a band at 1 kb. However, Shen Qian result was still not acceptable as we supposed to get at least some lanes with two bands.
Hong Ni decided that she would do the PCR to check the quality of the sample.
Today we also inoculated some bacteria. The bacteria is E.coli that had been transformed with plasmid vector with an insert. Each of us inoculated 8 colonies. After we finished with inoculation, Hong Ni briefed us on what is the schedule for tomorrow. But while she was explaining to us on what we were supposed to do on the next day, prof Hong came. Instead you were wondering, Hong Ni is professor daughter and she is currently doing her PhD in her father's lab.
He then told us that we were not supposed to be so attached to Hong Ni as she has her own project. Prof Hong then brought us to his office and tell us what we were supposed to be doing and gave us some home works. We were supposed to memorize at least 10 different restriction enzymes, their sizes, recognition sites and buffers. He also told us to practice opening and closing the 1.5 ml eppendorf tube with one hand. After that, we went to the shaker where we incubate our inoculation tubes as we thought that we need to loosen the cap.
At the shaker, Prof Hong found some inoculation tubes that were two third filled with LB medium. Luckily those were not belong to us. We are supposed to fill one fifth of the tubes as the bacteria needs air to grow.
After we had our dinner, we went back to the lab to see Hong Ni's PCR result but she also only get primer dimer bands. So, we were wondering what actually went wrong.
The first thing that we did in the morning when we arrived in the lab was to run a gel for our PCR (genotyping) result. We did not get the band that we wanted. Instead we got a single band for each lanes on uniform size, the size of the primer dimer, which means that our PCR result is not successful. Thus, Hong Ni asked us to redo our PCR.
We repeated every step, but this time we made our own PCR master mix, and run a gel again. At the end of the experiment, I still got primer dimer but Shen Qian got a band at 1 kb. However, Shen Qian result was still not acceptable as we supposed to get at least some lanes with two bands.
Hong Ni decided that she would do the PCR to check the quality of the sample.
Today we also inoculated some bacteria. The bacteria is E.coli that had been transformed with plasmid vector with an insert. Each of us inoculated 8 colonies. After we finished with inoculation, Hong Ni briefed us on what is the schedule for tomorrow. But while she was explaining to us on what we were supposed to do on the next day, prof Hong came. Instead you were wondering, Hong Ni is professor daughter and she is currently doing her PhD in her father's lab.
He then told us that we were not supposed to be so attached to Hong Ni as she has her own project. Prof Hong then brought us to his office and tell us what we were supposed to be doing and gave us some home works. We were supposed to memorize at least 10 different restriction enzymes, their sizes, recognition sites and buffers. He also told us to practice opening and closing the 1.5 ml eppendorf tube with one hand. After that, we went to the shaker where we incubate our inoculation tubes as we thought that we need to loosen the cap.
At the shaker, Prof Hong found some inoculation tubes that were two third filled with LB medium. Luckily those were not belong to us. We are supposed to fill one fifth of the tubes as the bacteria needs air to grow.
After we had our dinner, we went back to the lab to see Hong Ni's PCR result but she also only get primer dimer bands. So, we were wondering what actually went wrong.
Labels: Bacteria Inoculation, PCR
posted by Melinda Leonie @ 7:28 AM
0 Comments:
Post a Comment
Subscribe to Post Comments [Atom]
<< Home