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 :)

The Project

Hi there,

I thought i'd give a brief introduction to the project and why i'm doing it.

I'm working with Dr. Felicity Watts in the Sussex Centre for Genome Damage and Stability. I feel very lucky to have been given this placement by the Biochemical Society and the Genome Centre because it has given me a chance to experience what it is like to work in DNA stability and repair research which is what I believe I may want to do in the future.

The project I have been given to work on concerns the loss of heterozygosity (LOH) in chromosomes. This is where processes have caused mutations that cause a loss of function in both alleles of a gene. LOH often involves the removal of multiple genes and even whole chromatid arms. In diploid cells such as our own, we should have two copies of each gene and if one ceases to work the other can fill in for it. LOH removes this second copy making the likelyhood of cancer or other diseases much higher.

The centromere is extremely important in chromosome segregation and mutations occuring in the centromere repeats may be what causes this loss and chromosomal rearrangement. I will be studying the stability of the chromosome with mutations placed in different locations using a genetically engineered mini chromosome in the fission yeast; S. pombe.

Well thats all i'll say for now, lest my post become to essay-like. Should make a very interesting project though!