Selection of the optimal method for DNA extraction from big-seed false flax (Camelina sativa (L.) Crantz)
Year: 2024
Type: article
Abstract: To select a fast and reliable protocol for DNA extraction from Camelina sativa, six different methods were tested: CTAB (cetyltrimethylammonium bromide) 1% and 2%, DiamondDNA, Extran-3, D-Plants-5, and the MagnoPrime FITO kit, which allows DNA extraction on an automatic station. Sprouts and seeds of Camelina sativa were used as material. The quantity and quality of the obtained DNA samples were determined by spectrophotometry; three parameters were assessed: DNA concentration, protein contamination (А 260 / А 280 ), contamination with polysaccharides, fatty acids and other organic compounds (А 260 /А 230 ). In addition, PCR was performed with all samples to detect amplification. The mean value, standard deviation and confidence interval were calculated for the DNA concentration. When using the DiamondDNA and D-Plants-5 kits, a low DNA concentration (confidence interval) was observed – 5.31-15.43 ng/μl and 1.91-7.05 ng/μl, respectively, as well as the absence of amplification in all samples. The use of CTAB 1% and 2% ensured the isolation of samples with the highest DNA concentration: the maximum value of this indicator in the experiment was 136.38 ng/μl and 115.99 ng/μl, respectively. However, this method is labour-intensive, time-consuming and potentially toxic. The Extran-3 and MagnoPrime FITO kits were recognised as the most suitable for isolating DNA from camelina, with the DNA concentration (confidence interval) equal to 23.31-75.02 ng/μl and 33.55-97.51 ng/μl, respectively. Extran-3 allows to isolate the purest DNA (А 260 /А 280 – 1.96-2.02 units; А 260 /А 230 – 1.64-2.10 units), and MagnoPrime FITO, thanks to its operation on an automatic station, allows to isolate DNA from a large number of samples in a short time.
Language: English
Subfield: Molecular Biology
Domain: Life Sciences
SDG: Zero hunger
Open Access status: hybrid
OpenAlex ID: W7203749546