EMBL lambda replacement vectors: What insert size range can be accommodated by EMBL3 and EMBL4 lambda vectors?
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A6–7 kb
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B15–25 kb
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C40–44 kb
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D1–2 kb
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EGreater than 60 kb routinely
Answer
Correct Answer: 40–44 kb
Explanation
Introduction / Context:EMBL3 and EMBL4 are classic lambda replacement vectors designed to accept large DNA inserts by removing a central nonessential “stuffer” fragment. Knowing their insert capacity is important for planning genomic library construction and ensuring efficient packaging.
Given Data / Assumptions:
- Lambda packaging has size limits that define total recombinant genome length.
- Replacement vectors allow larger inserts than insertion vectors.
- EMBL3/EMBL4 are optimized for large genomic fragments.
Concept / Approach:By replacing a stuffer region, EMBL vectors can accept inserts on the order of tens of kilobases while keeping the total DNA within the λ packaging window. Their advertised capacity commonly falls in the ~40 kb range, enabling robust genomic library coverage.
Step-by-Step Solution:
Recognize vector type → replacement (large inserts).Match to typical capacity → approximately 40–44 kb inserts.Eliminate smaller ranges that fit insertion vectors or plasmids.Verification / Alternative check:Library construction manuals specify that EMBL replacement vectors handle inserts near 40 kb to obtain high-titer packaged phage.
Why Other Options Are Wrong:
6–7 kb or 1–2 kb: too small; typical of plasmid or insertion-vector scales.15–25 kb: under the capacity of EMBL replacement vectors.> 60 kb: exceeds λ packaging constraints.Common Pitfalls:Confusing total genome length with insert length; forgetting to account for the remaining λ arms when calculating packaging size.
Final Answer:40–44 kb.