Density-gradient centrifugation in cesium chloride: DNA labeled with heavy nitrogen (15N) will band where relative to DNA containing light nitrogen (14N)?

Difficulty: Easy

Correct Answer: Below the 14N DNA band

Explanation:


Introduction / Context:
CsCl density-gradient centrifugation separates macromolecules by buoyant density. Classic Meselson–Stahl experiments used 15N labeling to distinguish heavy vs light DNA and demonstrate semi-conservative replication. This question asks where 15N-labeled DNA bands relative to 14N DNA.


Given Data / Assumptions:

  • 15N increases DNA buoyant density compared to 14N.
  • In a stable CsCl gradient, denser species migrate to positions of higher density (farther from the axis of rotation).


Concept / Approach:
Heavier (higher density) molecules band closer to the bottom of the tube (i.e., farther out in a swinging-bucket rotor). Therefore, 15N-DNA bands below 14N-DNA.


Step-by-Step Solution:

1) Recall that buoyant density determines equilibrium position.2) 15N substitution increases DNA density.3) Thus, the 15N band forms lower than the 14N band in the gradient.


Verification / Alternative check:
Published density values show a measurable shift (~0.016 g/ml) between 15N and 14N DNA, yielding resolvable bands.


Why Other Options Are Wrong:

  • Above: would imply lower density, which is false for 15N DNA.
  • Co-incident: contradicts the density difference.
  • “All of these”: rotor speed influences resolution, not which one is denser.
  • Meniscus: banding occurs at the density-matching point, not at the surface.


Common Pitfalls:
Confusing tube orientation; in most diagrams, “below” denotes the higher-density region toward the outer radius.


Final Answer:
Below the 14N DNA band

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