All entries for Thursday 05 July 2007
July 05, 2007
After my rather upbeat first entry i can’t say i am as entusiastic now! I am looking at the distribution of methanogens in the guts of higher temites. presently i am working with two samples of DNA extracted frome the guts of termites; sample 001 and sample 133. At first i had to learn the biological tecniques needed to amplify the sections of DNA i was to look at.
The Polymerase Chain Reaction (PCR)
I have simplified the events of PCR so everyone can get a faint idea of what i’m doing below:
1. Heating the DNA to seperate the two strands.
2. Designed, archael specific, primers bind to both strands of the DNA.
3. Taq polymerase extends the primers to make copies of the archaeal specific DNA fragments.
I have found PCR relativley simple. As my supervisor says it is like cooking, as long as you follow the recipe it tends to work, however there has been occasions where i must have added a few ingredients of my own as it has not always been successful.
This technique seperates the PCR products by size creating bands of DNA that correspond to a marker that is always used in the first well.
At fist i was just carrying out simple PCR’s that were guaranteed to work, such as primers used on bacterial DNA. As these seemed to work fine i progressed to archaea DNA using non specific primers and the termite gut samples, some of these did not work as well and after much persiverance i acheived the results i wanted. I have now progressed onto termited group 1 specific archaeal primers – the particular group i am interested in and will let everyone know how it goes!