OBJECTIVES: The present research was conducted to assess incidences of root microcracks caused by hand and rotary file system at different lengths MATERIAL AND METHODS: This in-vitro study was undertaken to assess incidence of root microcracks caused by hand and rotary file system at different lengths In total, 100 the mandibular premolar with straight roots determined with intact, fully formed apices were taken. Samples were randomly distributed into 5 groups based on the file system used: a) Group A: Control, b) Group B: Reciproc, c) Group C: WaveOne, d) Group D: One Shape and e) Group E: ProTaper. Pearson Chi-square test was used to determine the differences between groups. The dentinal defects were expressed as percentage of samples with microcracks in each group. Level of statistical significance was set at p-value less than 0.05. RESULTS: The chi square test was used to compare the Distribution of the number of teeth in which cracks were observed on the horizontal sections. It was found to be significant with group ProTaper showing maximum cracks at 3,6,9 mm level as compared to other file systemCONCLUSION: Nickel-titanium instruments causes cracks on the apical root surface or in the root canal wall. ProTaper causes maximum dentinal cracks as compared to other file systems