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Background: Electromyographic (EMG) signals obtained from a contracted muscle contain valuable information on its activity and health status. Much of this information lies in motor unit potentials (MUPs) of its motor units (MUs), collected during the muscle contraction. Hence, accurate estimation of a MUP template for each MU is crucial. Objective: To investigate the possibility of improving MUP template estimation using the wavelet denoising technique. Material and Methods: In this analytical study, several MUP template estimators were developed by combining conventional estimation methods and wavelet denoising techniques. A MUP template was initially estimated using conventional methods such as mean, median, median-trimmed mean, or mode. Thereafter, it was post-processed using the wavelet denoising technique. Results: Evaluation results of the studied estimators using 40 simulated EMG signals with a true template for each constituent MUP train showed that augmented wavelet- based template estimation methods are more reliable than conventional methods. However, on average, wavelet denoising was not much effective. Around 40 MUPs of a MU is sufficient to estimate its MUP template. Conclusion: Although wavelet techniques are effective in EMG signal analysis, here wavelet denoising did not practically improve MUP template estimation. Considering computational simplicity and estimation error, the two methods median and median-trimmed mean are practical estimators that can provide a good estimation of a MUP template for a MU when approximately 40 MUPs are available. Nevertheless, the baseline noise level in the MUP templates estimated using the median-trimmed mean method is slightly lower than that in the templates estimated using the median method.