Question

Why sticky ended ligations are carried out at temperatures lower than room temperature?

(a) It is so because the vibrational and kinetic energy of the molecules at room temperature is lower than that of the energy required to break the bonds holding the ends

(b) The energy required to break the bonds holding the ends is very less than that of the kinetic and vibrational energy at room temperature

(c) The enzyme carrying out ligation is unstable at low temperature

(d) The sticky ends created, don’t just relegate at low temperature

I got this question during a job interview.

My doubt stems from Categories of Ligation Reaction in chapter Basics of Genetic Engineering of Genetic Engineering

Answer 1

Correct choice is (a) It is so because the vibrational and kinetic energy of the molecules at room temperature is lower than that of the energy required to break the bonds holding the ends

To explain I would say: The sticky ends are held together by the hydrogen bonds. These hydrogen bonds are weak in nature and the energy required to break them is very less than the kinetic and vibrational energy of the molecules at room temperature. Hence the reaction is carried at 4 degrees.