Practice Makes Perfect

 Gel Electrophoresis

    This week we focused on making gels, as well as adding samples to the gels. The goal behind it was to get used to putting samples inside the gels since they can easily be penetrated if not careful. Gel electrophoresis is important in the lab setting for separating DNA, RNA, or proteins. It uses an electrical charge to pull the samples through the gel, and once your sample has ran through the gel, it is placed over a UV light and the bands are read. Compared with a "ladder", you can determine the size of your samples base pairs. We originally started with practice gels, since they are rubber it takes a lot more to puncture the surface. We added our ladders as well as samples to the practice gels. 

This picture is the practice gel sitting inside of the the chamber. We have 2 "samples" in the middle, and 2 "ladders" on the outside. As you can see from the photos there is air bubbles inside the ladder wells (we do not want that).



    After everyone was comfortable loading the samples and ladders into the wells, it was time to make our own gels. We started with 120mL of TAE and we are making a 1% solution. So we take our 120mL and 1% of that, which is 1.2 grams, meaning for 120mL of TAE we add 1.2 grams of agarose powder. We heated the solution for one minute intervals, making sure to take the solution out and mix it if it began to boil. After the liquid is clear you let it sit for around 5 minutes to cool, and once cool you place inside the chamber. Once the solution is in the chamber, allow it to sit for around 20 minutes so it will coagulate. 

These photos are of our gel solution, as well as the gel cooled down and coagulated. 



    Once the gel has cooled and the dams and comb are removed from it, to stop the gel from drying out, we add our 1 times TAE solution to the chamber. Submerging the gel entirely. After the gel is submerged, we can add our samples as well as ladders. 

These photos are of the gel inside the chamber, with our "sample" and "ladder". 





    Since this week was for practice, we did not run the gels with electricity, or place them over UV lighting. For the sake of practicing, our "samples" were 10uL of water, and 2uL of dye, and the "ladders" were 4uL of water, and 2uL of dye. Next week we will be doing real gels and running our plasmid that we extracted. Everyday that goes by I find myself feeling more and more comfortable in the lab setting, which could be a good thing, or a potentially bad thing. 

Comments

  1. EricaMarch 12, 2022 at 11:20 AM

    Haha, good observation-- comfort can swing both ways. Nice work!

    ReplyDelete

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