A comparative evaluation of DNA yield, purity and integrity from different body segments of Apis mellifera using phenol–chloroform and commercial extraction

Efficient DNA extraction is essential for molecular studies in honey bees, yet extraction success may vary depending on methodology and tissue type. This study compared a classical phenol–chloroform protocol and a silica membrane–based commercial kit for genomic DNA isolation from head, thorax and abdomen tissues of the honey bee (Apis mellifera). DNA integrity, yield and spectrophotometric purity (A260/A280) were evaluated under standardized laboratory conditions. Agarose gel electrophoresis revealed that phenol–chloroform extraction produced high-molecular-weight DNA with minimal degradation across all body segments, whereas the commercial kit yielded weaker bands and partial smearing. Quantitative analyses showed that phenol–chloroform extraction generated significantly higher DNA concentrations than the commercial kit in all tissues. Two-way ANOVA confirmed a highly significant effect of extraction method on DNA yield (p < 0.001), while body segment alone did not significantly influence yield. However, a significant interaction indicated segment-dependent variation in extraction efficiency. DNA purity also differed significantly between methods (p < 0.001), with phenol–chloroform extraction producing more consistent A260/A280 ratios, particularly in thorax and abdomen samples. Overall, the extraction methodology was the primary determinant of DNA yield and quality. Although commercial kits offer advantages in speed and convenience, phenol–chloroform extraction remains a reliable method for obtaining high-quality genomic DNA suitable for molecular studies in A. mellifera.