Kinetics of PCR product formation: After the 4th cycle of PCR, how many double-stranded DNA molecules comprise precisely the target-length amplicon (bounded by both primers)?
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ATwo ds DNA molecule
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BThree ds DNA molecule
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CEight ds DNA molecule
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DFour ds DNA molecule
Answer
Correct Answer: Four ds DNA molecule
Explanation
Introduction / Context:In early PCR cycles, products include over-extended strands. Target-length amplicons (bounded on both ends by primer termini) first appear after the second cycle and accumulate in a predictable way. Understanding this helps explain exponential amplification and band purity.
Given Data / Assumptions:
- Two primers flank the target.
- Cycle counting starts at the initial denaturation/extension sequence.
- We are counting only the exact-length double-stranded products.
Concept / Approach:Exact-length product appears starting in cycle 3 and then doubles each cycle thereafter. A classic rule: after cycle n, the number of precise-length ds molecules equals 2^(n-2) (for n ≥ 3). Thus cycle 3 yields 2, cycle 4 yields 4, and so on.
Step-by-Step Solution:At cycle 1 and 2: products include over-extended strands; no exact-length ds product at cycle 2 end.Cycle 3: first time both ends are primer-defined → 2 exact-length ds molecules.Cycle 4: doubling from cycle 3 → 4 exact-length ds molecules.
Verification / Alternative check:Derive using strand-tracking diagrams showing how primer-defined ends accumulate and then dominate the reaction in later cycles.
Why Other Options Are Wrong:
- Two (A) corresponds to cycle 3, not cycle 4.
- Three (B) does not follow doubling logic.
- Eight (C) corresponds to cycle 5 for exact-length molecules.
Common Pitfalls:Counting total DNA molecules instead of exact-length amplicons; totals include heterogeneous lengths early on.
Final Answer:Four ds DNA molecule