The mechanism of hydrogen abstraction in the reaction of 3-methyl-1-butanol with an OH radical was investigated by quantum chemical calculations. The total reaction consists of five channels (R1–R5), with channel R3 having the lowest energy barrier (5.4 kcal·mol−1) and channel R4 having the highest energy barrier (13.2 kcal·mol−1). All channels are exothermic reactions with lower energy barriers. Thermodynamic parameters, including constant pressure molar heat capacity CPo, enthalpy of formation ΔfHo, and entropy So, for all reactants and products, were researched using the CBS-QB3 level across a temperature scale of 298–5000 K. The harmonic and anharmonic rate constants of all reaction channels were calculated at the CCSD(T)/6-311++G(d,p) level for a temperature range of 223–4000 K. Throughout the temperature range, except for R2, the harmonic rate constant of all reaction channels is consistently higher than the anharmonic one, and it can be seen from the results that the anharmonic effect is significant and cannot be ignored. Meanwhile, the polynomial coefficients a1–a7 for the thermodynamic parameters and the polynomial coefficients A, n, and E for the kinetic parameters were fitted using the principle of least squares.