Sds Kit
Sds Kit
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How much is in a powder buffer?
I am doing a science fair experiment on testing the DNA of bananas and well I need alittle help. The materials list a powder buffer, but im doing everything on my own, so I didn't buy the kit online. So i need to know how much papain enzyme, baking soda, SLS/SDS (liquid soap), and salt.
So it would be totally awesome if someone could help me because this project is due in four weeks and i don't want to be doing this last minute. Plus the bananas are ready to be tested. if anyone knows about this project, and has done it before please feel free to answer. So please answer my plea like A.S.A.P.
Thanks a bunch of bananas!
You want to buffer Taq polymerase at pH 8.4. Ordinarily you'd use a tris (tris(hydroxymethyl)aminomethane) buffer (pKa = 8.0; H tris / tris). The closer the pKa of your buffer is to your target pH, the better the buffer is able to resist pH changes for a given concentration of buffer. The buffer pKa should be within about 1 pH unit of your target pH or the the buffer capacity will be too low to be of any use. Baking soda has a pKa of 10.4 (HCO3- / CO3--), so it's not a very good choice as a buffer at pH 8.4. A phosphate buffer (pKa = 7.2; H2PO4- / HPO4--) is closer, but will still be a very poor buffer at pH 8.4.
If you don't have access to tris, you might try using lysine or arginine as a buffer (pKa ~ 9.0). You can buy these amino acids as supplements at a health fool store (obscenely overpriced, of course). Mash 'em up with a mortar and pestle, dissolve 'em in water or slightly acidic (pH > 3-4) solution, filter out the solids leaving only dissolved amino acid (hopefully). You may want to let them react with the papain before raising the pH to 8.4 for the Taq (in case there are any nucleases lurking in the vitamin pills). If you do this, you'll want to prepare an AA buffer and test its buffer capacity before wasting your time and your Taq on a buffer that doesn't work. (Health store vitamins ain't always what they claim to be.)
You probably also know that you need some magnesium in the pot for the Taq to work. Prepackaged buffers include MgCl2, but any source of Mg++ will do.
So, if you're doing it all on your own, how did you obtain the Taq?
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