Abstract

GUTIERREZ-CONTRERAS, Cecilia Nataly; BARRAZA-SALAS, Marcelo; MERCEDES PORTH, Ilga  and  WEHENKEL, Christian. Improvements to a commercial DNA extraction method for high-quality nucleic acid extractions from Populus tremuloides Michx. vegetative buds. Polibotánica [online]. 2026, n.61, pp.205-220.  Epub May 18, 2026. ISSN 1405-2768.  https://doi.org/10.18387/polibotanica.61.12.

Quaking aspen (Populus tremuloides) is a tree species of interest due to the potential discovery of an adaptive history in its genome, which is why the correct isolation of its DNA is essential. In this study, two extraction methods based on the NucleoSpin™ Plant II kit were investigated using vegetative buds from eight paired samples. Method 1 used buffer PL1, a homogenizer, and a 30-minute incubation with RNase; Method 2 employed buffer PL2, glass beads, and a 45-minute incubation with RNase. DNA quality and quantity were assessed by spectrophotometry, gel electrophoresis, and PCR amplification. A paired t-test revealed that Method 1 yielded a significantly higher DNA concentration than Method 2 (mean difference = 71.98 ng/µL; 95% CI 17.3-126.7; p = 0.017; dz = 1.10). In contrast, no significant differences in the 260/280 ratio were observed, either by the paired t-test (mean difference = 0.10; 95% CI −0.08-0.28; p = 0.219) or by the Wilcoxon signed-rank test (median difference = +0.02; 95% CI −0.01-0.295; p = 0.25). Method 1 produced higher yields but with DNA fragmentation visible on the gel, which may interfere with some analyses. Method 2 showed a lower concentration but slightly clearer bands on the gel, indicating better DNA integrity. PCR was successful in both cases, but the higher DNA integrity in Method 2 suggests that this method is more suitable for studies requiring intact DNA. Although Method 1 yields a larger amount, Method 2 is preferable when DNA integrity is critical for genomic analysis.

Keywords : quaking aspen; isolation; PCR amplification; DNA integrity; genetics.

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