Hands-on plasmid construction guide

critical

Confirmed colony later shows recombination or deletion

Initial screening looked correct, but on re-verification the insert is missing pieces or the vector has rearranged.

💡 4 · ✓ 4

severe

Double digest band pattern does not match prediction

Picked colonies grow normally but the diagnostic double-digest yields bands of unexpected size on the gel.

💡 5 · ✓ 5

moderate

Star Activity Producing Extra Bands

Extra DNA bands appear on gel beyond expected digest fragments, indicating cleavage at non-canonical recognition sequences due to relaxed enzyme specificity.

💡 4 · ✓ 5