What is the function served by each ingredient in the fractionation buffer? You
ID: 478762 • Letter: W
Question
What is the function served by each ingredient in the fractionation buffer? You have just accomplished the feat of isolating nuclei from the ova of a "golden goose" The ideal condition for polluting the nuclei involves centrifuging at 770 g for 10 minutes in a rotor of radius 11.02 cm. At what speed (in rpm) was the sample centrifuged? Your friend in California wants to repeat your procedure, but he has a rotor of radius 17.2 cm. At what speed (in rpm) should he centrifuge to pellet the "golden nuclei"?Explanation / Answer
Q.1: The ingradients used in fractionation buffer are;
Salt: Salts like NaCl, KCl etc. are used to establish an ionic strength in the buffer solution.
EDTA- free complete tablet protease inhibitors are used to inhibit protease enzyme. This prevents the catabolism of protein by the hydrolysis of peptide bnd.
dithiothreitol (DTT) is used as a reducing agent.
Distilled water is used to adjust pH.
Buffers like HEPES and Tris etc. are sued as buffer.
Q.2:(a): Given radius, r = 11.02 cm = 11.02 cm x (1 m / 100 cm) = 0.1102 m
a = 770 g = 770 x 9.8 m/s2 = 7546 m/s2
The formulae for centripetal accleration of a centrifuge is
a = V2 / r
=> V = square root ( a x r)
=> V = square root(7546 m/s2 x 0.1102 m) = 28.84 m/s (answer)
(b): Given r = 17.2 cm = 0.172 m
In both cases the accleration should be same.
a = 770 g = 7546 m/s2 = V2 / r
=> 7546 m/s2 = V2 / 0.172 m
=> V = square root (7546 m/s2 x 0.172 m) = 36.03 m/s (answer)
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