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How to make an acrylamide gel for electrophoresis
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[Music] when placing the glass in the casting frame make sure it's on a flat surface so it'll seal tightly when placed in the casting stand [Music] you may want to Mark the glass as a guide so you know how much running gel to add ideally Your Mark should be about 1 cm below the teeth of the comb you can insert the comb to double check once the APS and tet are added the solution will start to polymerize so try to add them as quickly as possible [Music] to add the running gel solution place the tip of the PIP pet in the corner of the space between the short plate and the spacer plate it's a good idea to leave a little bit of solution in the pipet to avoid introducing bubbles into the gel continue filling until the solution reaches Your [Music] Mark adding isopropanol to the top of the gel will keep the solution from forming a meniscus within the glass resulting in a very smooth surface don't forget to remove the isopropanol layer before adding the stacking gel [Music] [Music] [Music] [Music] once the running gel solution has polymerized and formed a plug you may add the APS and Ted to the stacking gel solution and then add it to the top of the cassette typically you should allow 20 minutes for polymerizing [Music] [Music] ation be careful not to damage the gel while removing the isopropanol layer [Music] [Music] remember that APS and Ted cause the solution to polymerize so add them immediately proceeding addition to the cassette [Music] bring the level of the stacking gel solution to the top of the short glass [Music] when inserting the comb some of the stacking gel solution will spill over wait until the solution is polymerized before cleaning the acrylamide is considered toxic until polymerization [Music] occurs when a gelatinous plug forms the vile you are ready to remove the comb from the gel the wells can be a little tricky to find Once the comb has been removed from the finished gel so marking the first well prior to removing the comb can save you some time be careful not to remove the comb at an angle because this can damage the wells [Music] you are now ready to run your gel through electropheresis to examine your sample good luck [Music]
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