Making of the gel—This is not a very interesting or exciting part of research, but it needs doing. Like Jell-O needs to be mixed up and put in the fridge before you can enjoy your jigglers, same idea with these gels. The basics are that you start with acrylamide (careful, it’s a neurotoxin!) and then you cause it to polymerize (the “poly” part of the name). Polymerization solidifies the acrylamide, so while it’s still liquid you need to pour it between the glass plates (ie, Jell-O mold) to solidify there. Once that’s done, you do it again (sort of like making multi-layer Jell-O, but without the colors or flavors). This time you add a “comb” to the top so that you get notches in the top. (see figure, picture of gel from BioRad)
Once it is all solidified you are ready to put your cell samples into the notches you’ve made. Apply a little electrical current and the proteins will move through the matrix that you’ve created by polymerizing the acrylamide. The small proteins can navigate through the matrix faster than the big ones, so you can separate proteins based on size.
Anyway, all of that explanation is so that I can say that the other day I had a bad day with polyacrylamide gels. I went into work planning on running a gel that I had made the day before. One of the first things I did was break one of the glass plates. Oops. So I started to make some more. All was well. Except that somehow I’d messed up and they weren’t polymerizing. Oops, again. I think I had better luck today—after disposing of the failed gels, I ordered some ready-made ones.
7 comments:
Keep at it! It will all come together. Or may I say ... gel?
It was frustrating setbacks like you describe that forever made my lab grades lower than my lecture grades. Oh, how I remember the stress!
On the bright side, it doesn't sound like you lost any of your protein samples in the ruined gels.
I'm reading this, thinking, "Bless that girl . . . poor thing had to be my lab partner in chemistry class!" Remember that, Sonnet? Mr. Carpenter? Thanks for saving my life in that class. I'm glad you could figure it all out from his teaching bottom to top instead of top to bottom. I think he'd be proud of you and impressed with what you're doing now!
As you were explaining the process of making the gels I though "that sounds laborious. I'm surprised those smarty scientists haven't figured out a way to pay someone else to do that." And then, your last sentence restored my faith in science! Behold, the glory of ready made! (Although, is it just not the same when you buy it that way? Those prepackaged jello cups are kind of gross, but "home-made" jello is YUM!)
Yes, Boss, we're sometimes willing to pay someone else to do the grunt work. But we scientists also tend to be somewhat paranoid and picky. Although we *could* have some one else do it for us, we aren't always sure they will do it *right* (ie, our way), so we'd just as soon do it ourselves. And sometimes we want Jell-O with fruit, but we don't want bananas or carrot shavings in it. We want raspberries, but raspberries are too acidic and might affect the congealing of the gelatin... And we don't even really like *green* Jell-O, or orange for that matter. Only the red ones. So that's why sometimes we opt out of paying someone to do the mundane stuff. Because of course, nothing goes wrong and all the data is perfect when we do it ourselves! :)
Forget the Jell-O. Ice cream is better anyway.
Wow, Sonnet! That is fascinating, but does sound quite laborious. I'm glad that they do have the ready made stuff for you, just in case there's a situation like this!
I think I'm with Catherine on this though---skip the Jell-o!
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