For the EQ approach, we employ our TIES protocol (successfully used for RBFEs ) that involves performing ensemble simulation at each intermediate alchemical state followed by ensemble averaging of the bootstrapped energy derivatives so obtained. Such averaged energy derivatives are then integrated using the principles of stochastic calculus to get the final ∆G along with associated uncertainties using equation 2. In the case of the NEQ approach, we adapt TIES to obtain the uncertainties associated with predicted free energies. We use the maximum likelihood estimator to obtain a ∆G value using the forward and reverse work values corresponding to all equilibrium replicas collectively at each end-state.