Abstract

This study evaluated the methods for preserving and extracting total DNA from ticks of the family Ixodidae, which are important pathogen vectors. Different preservation techniques, including isopropyl alcohol and RNAlater®, and DNA extraction methods (HotSHOT, acetate/acetic acid, salting-out, and phenol-chloroform) were used. The effect of maceration on the process of obtaining total DNA was also evaluated. Significant differences were absent between the methods performed with and without maceration. Among the extraction techniques evaluated, the HotSHOT and phenol-chloroform methods stood out, with no significant difference between the two. However, as the HotSHOT technique without maceration is inexpensive and requires fewer steps, it was considered superior to the phenol- chloroform technique. Regarding preservation, greater DNA amplification was obtained with RNAlater® than with isopropyl alcohol and was more beneficial for maintaining DNA amplification even after exposure to different conditions (storage time at room temperature [24–30°C] or freezing). These results highlight the importance of adequate preservation for successful molecular research, demonstrating the efficiency of RNAlater®. Our findings emphasize the relevance of appropriate extraction and preservation techniques for molecular research on ticks and contribute to an understanding of the most effective methods in this context.

Keywords

extraction of genetic material from ixodids, RNA later, HotSHOT, conservation, molecular analysis