Improvements to a commercial DNA extraction method for high-quality nucleic acid extractions from Populus tremuloides Michx. vegetative buds
DOI:
https://doi.org/10.18387/polibotanica.60.12Palavras-chave:
populus tremuloides , isolation, DNA, PCR amplification, DNA integrityResumo
ABSTRACT
Populus tremuloides is a tree species of interest due to the potential discovery of an adaptive history in its genome, making the correct isolation of its DNA essential. This study examined two extraction methods based on the Nucleo Spin™ Plant II kit, using growth buds from eight samples. Method 1 used buffer PL1, a homogenizer, and a 15-minute incubation with RNase; method 2 employed buffer PL2, glass beads, and a 45-minute incubation with RNase. Afterwards, the DNA was amplified by PCR. Method 1 produced an average DNA concentration of 137.9 ng/µL and a 260/280 ratio of 1.8 but showed DNA fragmentation on gel, which can affect some analyses. Method 2, although with a lower concentration (65.9 ng/µL) and a 260/280 ratio of 1.77, presented a clear band on the gel, indicating better DNA integrity. PCR was successful in both cases, but the greater DNA integrity in method 2 suggests it is more suitable for studies requiring intact fragments. Although method 1 offers greater quantity and purity, method 2 is preferable when DNA integrity is critical for genomic analysis.
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