The current research work presents a detailed and thoroughly validated computational micromechanical modeling methodology to study the damage initiation and propagation in a uni-directional (UD) glass fiber-reinforced non-crimp fabric (NCF) composite ply. Under the applied transverse tension and compression loads, the effect of various microscale material and geometrical parameters on the ply level stress–strain behavior is studied. To this end, along with the distinctly modeled individual microscale constituents of the UD NCF composite ply (axial fibers, backing fibers, and matrix), the generated RVE (Representative Volume Element) model consists of manufacturing-induced defects and variabilities such as voids as well as the backing fiber’s out-of-plane waviness. The fiber/matrix interface failure behavior in the generated RVE model is simulated using cohesive zone formulations that follow bi-linear traction-separation law. Whereas the hydrostatic pressure-dependent non-linear stress–strain response followed by the fracture of the epoxy matrix is captured using the linear Drucker-Prager plasticity model coupled with the stress triaxiality-based failure criterion. The backing fibers stress–strain and failure behavior is modeled using the linear elastic and isotropic material model in conjunction with the maximum stress criterion.The proposed numerical methodology is thoroughly validated both qualitatively and quantitatively by conducting detailed experiments on a neat epoxy as well as at the composites laminate level. Upon comparing the experimental and computational results, the observed knee or slope change in the bi-linear stress–strain response of the UD NCF composite ply under transverse tension is attributed to the loss of the load-carrying ability of the axial fiber bundle due to fiber/matrix interface debonding followed by the ply splitting. Under the application of transverse compression load, the composite ply failure behavior is governed by the formation of a dominant matrix plastic shear band in the axial fiber bundles, which is in turn triggered by the fiber/matrix interface failure. Under the applied loads in the matrix-dominated direction, the presented research work provides a detailed insight into the microscale damage initiation and propagation in a UD NCF composite ply and its consequent influence on the macroscale stress–strain response.