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When can ligation efficiency be higher?

Answer and explanation

Correct answer: When compatible sticky ends are present

The governing concept is complementary-end pairing before enzymatic sealing. Compatible sticky ends contain complementary single-stranded overhangs, so they can hydrogen-bond with one another and temporarily hold the DNA fragments in the correct alignment. This increases the local concentration and correct orientation of the 3′-OH and 5′-phosphate groups, allowing ligase to form the phosphodiester bond more efficiently. Therefore option A is correct. Boiling usually denatures the protein enzyme, absence of an energy source prevents or reduces ligation chemistry, and incompatible ends cannot pair properly. Blunt ends may also be ligated, but they generally require more favourable conditions and are often less efficient than compatible sticky ends.

Related tags

Ligation EfficiencyCompatible Sticky EndsBase PairingDna LigaseLigase EnzymesPrinciples Of BiotechnologyBiotechnologyClass 11 Mcq

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