Separating restriction fragments: Which electrophoretic media are commonly used to separate DNA fragments from a restriction digest?

Difficulty: Easy

Correct Answer: both (a) and (b)

Explanation:


Introduction / Context:
After restriction digestion, DNA fragments are separated by size using electrophoresis. The choice of matrix depends on the fragment length range and resolution required.



Given Data / Assumptions:

  • Agarose gels are suited for hundreds to many thousands of base pairs.
  • Polyacrylamide gels offer higher resolution for small DNA fragments (tens to a few hundred base pairs).
  • We are considering standard laboratory practice.



Concept / Approach:
Agarose concentrates (e.g., 0.8–2%) are tuned to resolve larger fragments; polyacrylamide (PAGE) provides a tighter matrix for fine resolution of small fragments or single base differences (e.g., sequencing gels). Both are valid and widely used for restriction fragment analysis.



Step-by-Step Solution:
Match fragment size range to gel type: large → agarose, small/high-resolution → PAGE.Recognize that both media are standard for separating DNA.Select “both (a) and (b).”



Verification / Alternative check:
Protocols for plasmid mapping frequently use agarose; mutation detection or small fragment resolution often uses PAGE, confirming both are appropriate.



Why Other Options Are Wrong:

  • A or B alone is incomplete because both are used depending on the application.
  • None (D) contradicts routine practice.



Common Pitfalls:
Using agarose to resolve very small fragments can blur bands; choosing incorrect acrylamide percentage can impair resolution for the target size range.



Final Answer:
both (a) and (b)

Discussion & Comments

No comments yet. Be the first to comment!
Join Discussion