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Q.Meselson and Stahl carried out centrifugation in CsCl2 density gradient to separate : (A) DNA from RNA (B) DNA from protein (C) The normal DNA from 15N-DNA (D) DNA from tRNA

CBSECBSE Class XII Board 2020MCQ· 1mImportance★★★★★
✓ Free question

Meselson and Stahl used CsCl density gradient centrifugation to separate normal DNA (¹⁴N-DNA) from heavy DNA (¹⁵N-DNA), based on their difference in density.

The question is about a landmark experiment in molecular biology — one that settled a long-standing debate on how DNA replicates. To understand what Meselson and Stahl were actually separating, you need to first see the problem they were trying to solve.

By the 1950s, scientists knew DNA was the genetic material, but they didn't agree on how it copied itself. Three models were on the table: conservative replication (the old molecule stays intact, and an entirely new one is built), dispersive replication (the old molecule gets chopped up and its pieces are scattered into two new molecules), and semi-conservative replication (each new molecule gets one old strand and one new strand). The last one was Watson and Crick's proposal, but it needed proof.

Meselson and Stahl designed an elegant experiment using an isotope of nitrogen. Normal nitrogen is ¹⁴N, but they grew bacteria in a medium containing the heavier isotope ¹⁵N. After many generations, all the bacterial DNA became "heavy" — every strand was labelled with ¹⁵N. Then they switched the bacteria back to normal ¹⁴N medium and let the DNA replicate. The key question was: what happens to the density of the DNA after one round of replication, and then after two?

To answer that, they needed a way to tell heavy DNA apart from light DNA. This is where the CsCl density gradient centrifugation comes in.

Note

Cesium chloride (CsCl) is a salt that forms a dense solution. When spun at very high speed in an ultracentrifuge, the Cs⁺ and Cl⁻ ions settle into a smooth gradient — densest at the bottom, lightest at the top. DNA molecules move to the point in the tube where their own density matches that of the CsCl solution. This is called "equilibrium density gradient centrifugation."

So, when Meselson and Stahl placed their DNA samples into the CsCl gradient and spun them, the DNA bands formed at specific positions depending on its density. DNA made entirely of ¹⁵N (heavy) bands lower in the tube. DNA made entirely of ¹⁴N (light) bands higher up. And hybrid DNA — with one heavy strand and one light strand — bands exactly in between.

That is the core of the experiment. They were not separating DNA from RNA, or DNA from protein, or DNA from tRNA. Those were not the point. The entire purpose was to distinguish between normal DNA (¹⁴N-DNA) and heavy DNA (¹⁵N-DNA), and to track how the proportion of each changed over successive rounds of replication.

Important

The purpose of the CsCl density gradient step was specifically to separate ¹⁵N-labelled DNA from ¹⁴N-labelled DNA. The other options — separating DNA from RNA, protein, or tRNA — are not part of this experiment.

The results were beautifully clear. After one round of replication in ¹⁴N medium, all the DNA was of hybrid density — exactly halfway between heavy and light. This ruled out conservative replication (which would have given one heavy band and one light band). After two rounds, two bands appeared: one hybrid and one light. This ruled out dispersive replication (which would have given a single band of intermediate density that gradually shifted). Only semi-conservative replication matched the data.

So the answer to the question is straightforward: the centrifugation was used to separate the normal DNA from ¹⁵N-DNA, based purely on density differences.

✓Final answer

In short, Meselson and Stahl used CsCl density gradient centrifugation to separate normal DNA (¹⁴N-DNA) from heavy DNA (¹⁵N-DNA), allowing them to track the density of DNA over successive generations and prove that DNA replication is semi-conservative.

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