Gel Electrophoresis

Gel Electrophoresis Lab

Vials of DNA.

Intro:

In this lab, we were able to complete gel electrophoresis! In order to do this, we had to make our own agarose gel, fill the wells and different dyes and finally electrophoresis the gel to gather the data that we needed to complete the experiment. Gel electrophoresis tells how big the DNA fragment is and where the restriction enzymes cut open the plasmid. Down below will be a better explanation of our lab process :)

Like a giant pool of gel bathing... but with electricity.

Procedure:

 Throughout this lab, there was a lot of hand shivering, especially when trying to carefully place the DNA substance inside of the Gel wells. What we basically had to do was prep the gels for the electrophoresis part. Doing this by carefully taking a micro pipet and placing the liquids into wells. It was quite difficult because it was very easy to blow away all of the DNA if we had let out the DNA too fast. For the 1st well, we had the Lambda DNA - in order to help us determine how long the base pairs were of the DNA we were testing for. 

The DNA placing begins!

The next well, we left it blank to easily compare lines. 3rd well we places the DNA that contained the restriction enzyme "PstI" - the 4th well we placed the DNA that had the restriction enzymes "PstI" and "HpaI" - for the 5th well we had "PstI" and "SspI" - for the last well we had all three restriction enzymes. After placing the DNA in their respected wells, we turned on the electrophoresis and let the DNA run.

Analysis: 

Using all the data from out gel, we could actually be able to draw out the plasmid and figure out where the restriction enzymes were.

Reason why is because each band represents a cut that the restriction enzyme made. The more enzymes we put in the more cuts we got and the more bands we saw. Also, being able to measure each band using our maker lane helped us determine how long each band was in terms of base pair size. 

Conclusion:

 This lab, for us, felt like a lab that real scientists would preform. This is something forensic scientists and DNA analysts do everyday. Also, for our group specifically, we feel achieved the best results. At least the most clear and accurate results. Below is the final picture of our gel. The bands stopped there because that is how fast they could travel within the amount of time we let it run in correlation to how large the DNA was. Each band represents an amount of base pairs. And so for the bands closer to the top represent the slower/longer DNA pieces. Opposite for the ones on the bottom. Also, the reason for the amount of bands is because we got to see where each restriction enzyme cut and how long each piece cut was. We enjoyed this lab very much.

Our final results.

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!