Question

Regarding point mutations observed in plasmid construction via Gibson Assembly, what might contribute to this issue?

A) PCR-induced mutations might arise due to polymerase errors, influencing the fidelity of DNA amplification.
B) DNA purification protocols could introduce contaminants leading to point mutations during plasmid construction.
C) Longer primers with increased melting temperatures are more likely to induce point mutations during PCR amplification.
D) Gibson Assembly itself causes random point mutations along the plasmid sequence, irrespective of PCR reactions.

Answer 1

Point mutations observed in plasmid construction via Gibson Assembly can be caused by PCR-induced mutations and contaminants introduced during DNA purification protocols.

Step-by-step explanation:

Point mutations observed in plasmid construction via Gibson Assembly can be caused by several factors. One possible contributing factor is PCR-induced mutations that might arise due to errors made by DNA polymerase during DNA amplification.

These errors can lead to changes in the nucleotide sequence of the plasmid. Another factor that can contribute to point mutations is the introduction of contaminants during DNA purification protocols. Contaminants may alter the DNA sequence and result in mutations. However, neither Gibson Assembly nor longer primers with increased melting temperatures directly cause random point mutations along the plasmid sequence. These options are not the primary contributors to the issue.