Today I was pleased to see that over the weekend a lot more colonies had grown on my antibiotic plates.
The colonies I'd picked the previous week however hadnt grown very well. My supervisor Felcity said that the colonies that the colonies that develop later on the plate are usually more likley to be what we are looking for. Especially as one of the first colonies to spring up was a contaminent.
As there were lots of colonies I spent a while picking more colonies to be spread on new plates. I made three plates with 25 colonies on each. Hopefully i'll have more success with some of those in future. There will be more chance of finding the right colony with 75 of them than with just the 4.
I was given some pipettes of my own to use. I tested them to make sure they were accurate by weighing the contents of their tips and checking it was the correct weight for the amount taken in.
After that, I begun to look into the next steps of the experiment as there was such a large amount of colonies growing and so more chance of finding the yeast colonies I want this time. I will be doing fluctuation tests on the yeast with the changed marker in comparison with the wild type yeast. Fluctuation tests as I said the other day are a measure of the rate of spontaneous mutation in bacteria. I will be running these to see whether there is a difference between the wild type and the chromosome with the new marker. This should then give way to more tests involving the creation of DNA "plugs" (DNA of the cells enacsed in agarose) which we may be able to use to show the exact form of the transformed chromosome to see if everything is where it should be. If everything is where it should be then the yeast strain is ready to use in chromosome observation experiments.
The fluctuation tests involve some statistics and will take a reasonable amount of time to carry out so I plan to read more about them so I know exactly what im doing. Having to repeat fluctuation tests if it goes wrong could take weeks.
Tommorow I will be going in early to watch how to make DNA plugs!
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