Neuro muscular Junction connects the nervous system to the muscular system via synapses between efferent nerve fibers and muscle fibers, also known as muscle cells. As an action potential reaches the end of a motor neuron, voltage-dependent calcium channels open allowing calcium to enter the neuron. Calcium binds to sensor proteins on synaptic vesicles called synaptobrevin triggering vesicle fusion with plasma membrane and subsequent neurotransmitter release from the motor neuron into the synaptic cleft. In vertebrates, motor neurons release acetylcholine (ACh), a small molecule neurotransmitter, which diffuses through the synapse and binds nicotinic acetylcholine receptors (nAChRs) on the plasma membrane of the muscle fiber, also known as the sarcolemma. nAChRs are ionotropic, meaning they serve as ligand gated ion channels. The binding of ACh to the receptor can depolarize the muscle fiber, causing a cascade that eventually results in muscle contraction. Neuromuscular junction diseases can be of genetic and autoimmune origin. Genetic disorders, such as Duchenne muscular dystrophy, can arise from mutated structural proteins that comprise the neuromuscular junction, whereas autoimmune diseases, such as myasthenia gravis, occur when antibodies are produced against nicotinic acetylcholine receptors on the sarcolemma. Disorders of the neuromuscular junction have a wide range of clinical presentations, which frequently poses a diagnostic challenge to evaluating clinicians. This article describes the tests used in the diagnosis of diseases of the neuromuscular junction, reviews the evidence supporting the use of each test, and proposes guidelines for their efficient utilization.