Gel Electrophoresis

 Gel Electrophoresis

    This week was a caveat from the previous weeks practice. The week prior we used practice gels to get our technique down for loading the wells. The practice gels were a lot more rigid compared to the actual gels used for the electrophoresis process. The basics of why we use gel electrophoresis is charging the molecules which then moves with the electric current through the gel, then verifying the base pairs within the sample used. We began by filling our chamber with 1x TAE as our buffer. For the samples we used 5 micro liters of DNA, 5 micro liters of water, and then 2 micro liters of loading dye. For the ladders we used 6 micro liters of the ladder DNA. Since this was also practice, a few of use shared gels. Once the gels were loaded with our samples and connected to our transformer, we began the process. For the transformer wee had it set to 85 mA, verified it was bubbling from both terminals, and waited for the samples to hit the back of the directional arrow. 

This first image is of our gels loaded, my group had some miscommunication and loaded our ladders incorrectly.


These two images are of the samples going through the gels. The dark blue is the wells, the purple is the samples going through the gels. 



Once the electrocution process was over, we placed the gels over UV light to view the ladders and samples. These two gels were not loaded with the correct ladders, that is why there is no ladder between them. 


As you can see in one of these images, the ladder is present between the two samples. In the other gel, the ladder was loaded as well, but someone punctured one of the wells and the dye spread throughout the gel, making it hard to read. 



    Another great practical to get more experience doing gels. From this we learned there's a specific ladder sample that needs to be loaded into the ladder wells, not the DNA being tested. We also got to see what happens when someone punctures a well and the DNA spreads throughout the gel. 

Comments

  1. EricaMarch 12, 2022 at 11:22 AM

    Oops! These things do happen, luckily you won't likely make that mistake again.

    ReplyDelete

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