How to Make an SDS-PAGE gel

today I'll be showing you how to cast an sds-page gel here it is the apparatus for casting the gel there are four different parts to this jaw paratus first of all you have two glass plates you have a short plate and then you have a toe plate then you also have a casting frame and a casting stand what you do here right now is you want to place the short plate in front of the toe plate like that and then put it into the casting frame after you plating the casting frame you secure it by clamping it and then you want to check that the bottom is flat where everything is done okay you can put it on the casting stand okay after we have assemble the jaw paratus what we need to do right now is we will take the comb and we'll place it in between the glass plates like that and we'll take a marker and we'll make a mark up approximately one centimeter beneath below the comb there we take the combat what we want to do right now is we want to add a little bit of water to the glass plate to see whether they are leaky or not so as you can see right now dr. adage is not leaky so therefore would rot out of the water using a piece of filter paper okay now we'll be casting the resolving Joe so I have added everything and the recipe is set for teammate teammate is going to initialize the polymerization reaction so after you add a teammate you should cast a job right away so now I have just added teammate to my solution so we want to pour the gel right away we want to pour the gel till the gel is at the same level as the mark it's okay if you introduce any air bubbles right now we'll fix that later so we'll pour the gel to the mark and then we'll remove the bubbles by adding a layer of isopropanol on top of the gel like that so after the resolving gel has polymerized will dry up the isopropanol using the filter paper again after we draw the isopropanol we can test the stacking gel so right now I'm going to have team it to the stocking Joe so we'll filled up the wrist of the plate restocking Joe we'll fill it all the way to the top like that right now you don't want to introduce any air bubble because you just going to get trapped and then we'll put in the cone and that's how you cast an sds-page gel

Channel: labtricks

Views: 536,325

Rating: 4.9281931 out of 5

Keywords: labtricks, science, lab, tips, tricks, techniques, SDS, PAGE, sodium, dodecyl, polyacrylamide, gel, protein gel, electrophoresis, casting, biochemistry, molecular, biology, chemistry, protein, video, tutorials

Id: EDi_n_0NiF4

Length: 4min 49sec (289 seconds)

Published: Mon Jan 18 2010