The current animal research investigation emphasizes animal wellbeing. Therefore, the present study recommends using non-invasive sampling procedures in scientific studies. The purpose of this study was to evaluate DNA extracted from 7 different parts of the feather (calamus tip, rachis (I-III) and barbs (I-III)) for sex identification using polymerase chain reaction (PCR) amplification. The study results showed that the calamus tip had the highest DNA concentration. DNA extracted from rachis and barbs can also be amplified and use for sex determination. Extracted DNA from the calamus tip, rachis (I), and barbs (I) showed accurate results for amplified PCR products when compared to the sex of known samples. The findings revealed that, in addition to the calamus tip, the rachis and barbs on the lower part of the feather surrounding on the calamus can be used to determine sex. Calamus tip collection by plucking is an invasive technique that could result in contusion and infection. The non-invasive sample collection method of cutting a portion of the feather is one option for supporting animal welfare guidelines. Furthermore, the non-invasive method presented in the study can be used to collect samples in other branches of molecular biology. Manuscript profile