pGLO Transformation Lab

 

Results from our lab. The four plates above are the +LB/amp,+ LB/amp/ara,-LB/amp, and the -LB.
The +LB/amp was the antibotic, the + LB/amp/ara was the arabinose sugar, and the negative plates were the plasmids.

 



This lab that we did in class was about how we collected information about the +pGLO cells with E coli grown on the plates. The plates that I would expect to find bacteria like the original non-transformed E.coli colonies that were intially observed would be on the LB plate because you can hopefully see everything growing because it is cabale of doing it. The LB/amp/ara plates would genetically have transformed bacterial cells because it would have had to take the DNA from the Amp in order to survive. The -pGLO and + pGLO plates should be compared to dertermine if any genetic transformation has occured because it would tell you if the plasmid transformed them. The negative plates were the control plates.

Observations that we collected was how there was alot of bacterial colonies on the + LB/amp, and there was around 35 colonies. The bacteria is a fluroscent green color. The bacteria in this plate didn't glow. On the + LB/amp/ara there was a less amount of bacterial colonies, and there was around 25 colonies. The bacteria was also a fluroscent green color. The spots on this plate glowed because it got a signal from the environment. The - LB/amp didn't have any spots, it was just a smear which was a fluroscent green color. Nothing occured on the - LB plate because there was just condensation on the plate. The traits that was originally observed for E.coli that was not altered was the colony shape and color. Traits that were different after performing the transformation procedure was the colony shape because there was a less amount of dots on the two positve plates, and on the -LB/amp the shape was totally different because there wasn't any dots. If the genetically transformed cells were acquired the ability to live in the presence of the antibiotic ampicillin then the other genes on the plasmid would have no bacteria growing. The changes that occured were due to the procedure that was performed because the + LB/amp didn't glow due to that glowing will only happen in the presence of arabinose. With the plasmid, the genes that code for the enzyme that digest arabinose are replaced with the GFP. So, without arabinose the gene is no longer working so that is why it didn't glow. In conclusion, I learned alot of neat information by doing this lab, and the transformation about pGLO cells with Ecoli grown on the plates.