PURPOSE The purpose of this study was to compare anterior cruciate ligament (ACL) graft length and tension throughout knee range of motion with transtibial, anteromedial (AM) portal, and all-epiphyseal drilling techniques with suspensory and apical femoral fixation. METHODS The three different femoral tunnel drilling techniques using the same intra-articular starting point within the center of the femoral footprint were performed on fresh-frozen cadaveric specimens. All groups underwent standard tibial drilling in the center of the ACL tibial footprint. FiberWire (Arthrex Inc., Naples, FL) was used to simulate anatomic single bundle reconstructions. Changes in graft length and tension were measured at knee flexion angles of 0, 30, 60, 90, 120, and 135 degrees. RESULTS Graft length and tension decreased from 0 through 60 degrees and subsequently increased from 90 to 135 degrees for all groups. The transtibial, AM portal suspensory, and apical fixation groups were similar. However, the all-epiphyseal tunnel with suspensory fixation had a significantly increased change in length (90, 120, and 135 degrees) and tension (120 and 135 degrees). CONCLUSION Transtibial and AM portal suspensory fixation and apical fixation demonstrate similar changes in length and tension throughout knee range of motion. The all-epiphyseal tunnel with suspensory fixation was associated with greater length and tension changes at higher degrees of knee flexion. All techniques demonstrated decreased graft length and tension with knee flexion to 60 degrees after which they increased with further knee flexion. CLINICAL RELEVANCE ACL graft length and tension change throughout knee range of motion and also depend on femoral tunnel orientation and fixation type. The use of an all-epiphyseal tunnel with suspensory fixation should be studied further for evidence of graft elongation.