My first day

Hi again,

Now that i'm done telling you about my project I can tell you about my first day at work! I spent the first part of the day becoming familiar with the labs, people, where things were and what the project itself was about.

My first aim in this project is to change the selective marker in the strain of S.Pombe I will be using later on.

Today I was shown how to use the polymerase chain reaction (PCR) procedure to amplify the new marker needed to replace the old one. A large concentration of the marker is needed to give a greater chance of transformation in the yeast culture. I was shown how to create a working PCR mixture and how to use a nice PCR machine to carry out the temperature change steps for the type of polymerase being used.

I got to make an agarose gel for later use in gel electrophoresis. I made this so that I could tell whether the PCR had been successful. I ran the PCR product next to a lamda Ecori/HINDIII marker, this marker is used like a ruler so that the size of the PCR product can be determined by comparison.

I then was able to view the results of the gel electrophoresis under UV light and take the picture below! :D This picture confirmed that my PCR was successful as I got the correct size fragments at 1.5kb:



Lastly I was taught how to grow a culture of the wild type yeast in preparation for tommrows transformation!

I really really enjoyed today, i feel I have learned alot and it was really nice to have a good result on the first day, hopefully i'll keep it up. Though i'm sure when it comes to doing it all by myself it will be rather challenging!

Thats all for now :)