pGlo Lab

pGLO Lab

Introduction:

 In the the pGLO lab, we were able to experiment with the transformation of genes. Genetic transformation simply means that we inserted an alternative gene to change the existing one. In this case, we transformed e-coli to become antibiotic resistant along with being able to glow in the dark. In order for this to work, we had to use a plasmid that carried the GFP gene we were looking for, transformation solutions and using heat-shock procedure. 

Procedure:

  The first step in this experiment was to label our micro test tubes as +pGlO and -pGlO. We then had to transfer an equal amount of our transformation solution (CaCl2) to each tube. After placing our tubes on ice, we added a portion of the bacteria colony to each tube. Next, we added plasmid to only the +pGlo tube. While our tubes were taking an ice bath, we prepared our nutrient agar plates by labeling them as shown below. After this, our tubes were ready for the heat shock. We first put them in ice, then in a warm bath for exactly 59 seconds, and then finally in ice once more. Next, we added some LB broth to each micro test tube. We then added the matching solution in the tube to the agar plate and spread it around. After taping the plates together, we left them to incubate overnight. In the morning, we took out our plates, and could already see adifference. There was a much larger amount of E. coli in the agar plate than there were the night before. Finally, it was the moment we had all been waiting for. We turned off all the lights, shined the UV light, and the E. coli glowed!!!

Analysis:

 The results boiled down are pretty simple. We learned a lot about how transformation worked with genes. We got curious about a few things. Like how come there wasn't as much growth on the AmpR plates as there were on the normal control? Come on, there are resistant are there? Well we seem to think that maybe not all of the ecoli were able to absorb and take in that plasmid we were providing. Thus killing off the ones who could not take in the gene. Where on the control, there weren't many problems for the ecoli.

Conclusion:

Luckily in this lab, no one suffered from any major E. coli infections. However, E. coli was harmed in the making of this lab. This was probably one of the first labs that our group felt like real scientists. This is because we successfully transformed the DNA of the E. coli using the plasmids we were given. As the pictures in our post show, we were one of the most successful groups in our period. Thats because we're cooler than absolute zero.

LET IT GLOOOW! LET IT GLOOOOOOW!