Introduction: The aim of this in vitro study was to evaluate the effects of different root canal instrumentation techniques and preparation tapers on the amount of apically extruded bacteria. Methods: The root canals of 98 extracted human mandibular incisors were contaminated with Enterococcus faecalis suspension. After incubation at 37 degrees C for 24 hours, the root canals were instrumented with K3 rotary files in a crown-down (CD) or full-length linear instrumentation technique (FL) by using 3 different root canal tapers (0.02, 0.04, and 0.06). During instrumentation, apically extruded bacteria were collected into vials containing saline solution. The microbiological samples were taken from the vials and incubated in brain-heart agar medium for 24 hours, and the numbers of colony-forming units (CFUs) were determined. The obtained. results were analyzed with t test and one-way analysis of variance for the comparisons between the instrumentation techniques (CD and FL) and the preparation tapers (0.02, 0.04, and 0.06), respectively. Tukey honestly significant difference test was used for pairwise comparisons. Results: The preparation taper had no effect on the number of CFUs when a FL instrumentation technique was used (P > .05). There was a statistically significant difference in the CFUs between FL and CD techniques when the preparation taper was 0.02 (P < .05). There was no statistically significant difference between the 0.04 and 0.06 preparation tapers in any of the instrumentation techniques (P > .05). Conclusions: Using a 0.02 taper in a CD manner results in the least amount of bacterial extrusion. The instrumentation technique did not seem to affect the amount of bacterial extrusion when 0.04 and 0.06 taper instruments were used for cleaning and shaping the root canal space.