So the objective of this week was to successfully find a vector that would be suitable for our experiment. We started by using two different vectors: HSA and HSA 4C. Along with trying to find the correct vector, we also wanted to see which restriction enzyme gave us the smoothest and clearest cut. We created six solutions and tested them by running them under a gel. We discovered that the best vector we would use is HSA and the best restriction enzyme would be Xho I. Finding the correct vector was an extremely tedious process which required many digestions. One aspect I truly dislike about working at the lab is all the waiting time. In all these processes such as Gel Electrophoresis and Digestions I have to wait about 45 minutes- 2 hours; I wish this process could be expedited. After using this gel, we extracted the best one using a Gel- Extraction Kit. I have used many kits this summer, either to purify or extract DNA, and I am fascinated how through reactions that the DNA can be cleaned and purified. Along with this, we were still working to find the best sample of insert DNA. Although, we found the correct size of the fragments (thanks to gels) we are noticing that not enough DNA is showing up after the PCRs. We continued to try and find a solution to this problem. By the end of the week, the DNA was still not as strong as we hoped but we decided to continue with our project, because I don’t have a lot of time left!
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