Mayen Dada's blog

This week has been busy, however i was able to find the correct primers to use in the PCR reaction mixture.  The colonies that i had from my  transformation are not working, so i have been busy repeating the transformations along with my left over A+B ligation mixture.  Hopefully, this experiment works by this weekend so i can move to the next step.  At the moment, my goal is to get my strain sequenced by next week and pratice other lab techniques.

I completed the lacZ assay and was able to make a few graphs on excel.  The result is closer to what it is suppose to be.  This week into the next week i will be focusing on purifing my nasD fragment and ligate it into the pTK-lac vector.  I am hoping that this procedure works better than my experience with the other experiments. So far, i have been learning how to make PCR reactions, operate the PCR machine and running my product on agarose and low-melting gels. 

This week i have been working on a lacZ Assay, which is a three day experiment.  The first part is 5hours and the other parts are about three hours.   The first time i began this assay i made a mistake at the 4th hour of the first part and had to repeat the experiment.   In this experiment i  am working with two strains the LAB2854 and ORB6188.  The purpose of this assay is to determine B-galactosidase activity in the bacteria during anaerobic bacteria, however  the experiment is still in progress.

This week we read a few jounal articles that had to do with finding where the NsrR binds and how this site would be affected if you have site-directed mutagenesis.  At the moment i am still learning and researching the different genes that are controlled by NsrR during anaerobic respiration.   Also, the professor went over various concentration concepts with dilutions and molarity.