Comparison of DNA extraction methods from Halocnemum strabilaceum (Amaranthaceae)
DOI:
https://doi.org/10.5281/zenodo.6498963Keywords:
Agarose Gel Electrophoresis, CTAB, PCR, Nano-Drop, plant tissueAbstract
Molecular techniques such as DNA extraction and DNA sequencing are playing an important role in studying the genetic makeup of the plant and identifying the evolutionary relationship using DNA barcoding. Extraction of DNA from plant tissue is often problematic, as many plants contain high levels of secondary metabolites that can interfere with downstream applications, such as PCR. Removal of these secondary metabolites usually requires further purification of the DNA using organic solvents or other toxic substances. In this study, we have focused on the DNA isolation process using three isolation techniques: the cetyl trimethyl ammonium bromide (CTAB) method that uses the ionic detergent hexadecyltrimethyl ammonium bromide and chloroformisoamyl alcohol, BioFACT, and GeneAll methods on desert/rangeland plant including Halocnemum. Quantity and quality of extracted genomic DNAs were compared by employing the spectrophotometer, Nano-Drop, agarose gel electrophoresis, polymerase chain reaction (PCR) methods, and molecular markers such as ISSR. Our results showed that the modified method of CTAB provided the best results than the BioFACT and GeneAll methods for extracting DNA from tissues of Halocnemum. We present a safe and cost-efficient DNA purification procedure and recommend using this CTAB method to extract DNA from plant tissues and to use the young leaf for the highest DNA yields.
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