How to Make and Load a Gel

    This week in lab, I learned how to make and load a gel. For this blog, I will be discussing the protocol on how to make a 30mL gel. Before starting the process of making my gel, I had to figure out what percent of agarose we should use. For my practice gel, I did 1% agarose (percentage determined by how small or large the DNA is that you're using). For my conversion, I did the following equation:

30mL  *  0.01  *  1000  =  300mg

            (percent  (conversion

             agarose)    to mg)

    I measured 30mL of TAE buffer in a flask a little over twice the volume. I then measured 300mg of agarose on my scale, added it to my buffer, then swirled until cloudy. Before putting my gel solution into the microwave, I made a stopper out of paper towel. After inserting the stopper, I put the solution in the microwave at 30 second increments until it was boiling. Once boiling, I let it sit for 5 minutes to cool down. As it was cooling, I set up my electrophoresis machine. I set up the electrophoresis by inserting my gel tray then my barriers. I ensured that they were tight to prevent leaks, then I added the comb. When my solution cooled after 5 minutes, I removed my stopper, swirled my flask, and poured it slowly and evenly into the gel tray. After pouring I checked for leaks, then quickly rinsed my flashed. I let the gel sit until hardened. Once my gel was hardened, I removed the barriers and poured TAE over it. I made sure to your it over the wells (to avoid air bubbles) then into the buffer wells. 

    I then proceeded to load my "DNA" (A mixture of just PCR water and dye for practice). The typical recipe used for gels is 5µL DNA, 5µL PCR water, and 2µL dye. Then of course we did 6µL of a 1kb ladder. When loading a gel, it is important to avoid piercing the sides and bottom with the micro pipette. Thats why when you load a gel, you want to load it by first making sure you're in the well and only being halfway into it. When loading my gel, I discovered that if you gently bump the sides of the well with the tip of your pipette, it is easier to gauge whether you're far enough into the gel or not. Due to the sample not having DNA in it, we did not run the gel.