Q.86
Which of the following is a limitation with chain termination sequencing?
EXPLANATION
Next-Generation Sequencing (NGS): Illumina Sequencing-by-Synthesis, PacBio & Oxford Nanopore
- Limitations of Sanger Chain Termination Sequencing:
- Intrastrand Secondary Structures: High GC content or inverted repeats within the template DNA can form stable hairpins and secondary structures.
- Polymerase Pausing: These structures physically impede DNA polymerase progression, causing premature termination ("stop bands" across all four lanes).
- Compressions in Gel Electrophoresis: Incomplete denaturation of secondary structures alters electrophoretic mobility, causing multiple fragments to migrate anomalously at the same position (gel compression).
Correct option: (3) Intrastrand base pairing can block the progress of the DNA polymerase and may also affect the migration of the molecules during electrophoresis
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