A Rapid, Economical and Hazardous Organic Solvent Free Method for DNA Extraction from Cassava
DOI:
https://doi.org/10.14456/thaidoa-agres.2021.16Keywords:
DNA extraction, cassava, interfering substance, sodium dodecyl sulfate (SDS), polyvinylpyrrolidone (PVP)Abstract
The extraction of good quality DNA from leaves of cassava is a complicated process
due to the presence of interfering substances such as polyphenols and polysaccharides that
affect the quality of DNA. In hexadecyltrimethylammonium bromide or CTAB method which
is widelyusedfor DNA extraction,hazardousorganic solvents suchasphenol andchloroform
are required to remove these interfering substances. The whole process does take
approximately 130 minutes making it unsuitable for extracting large number of DNA samples.
The objective of this research was to develop a DNA extraction method from cassava leaves
which is rapid, free of harmful organic solvents, does not use liquid nitrogen, as well as cost
and time effective. The DNA extraction method developed in this research was based on
sodium dodecyl sulfate/sodium chloride or SDS/NaCl method. The SDS/NaCl+PVP method
yielded DNA amount in the range of 1,431.4 - 1,510.3 µg per 1g of leaf sample with absorption
ratios A260/A280 of 1.74 - 1.90. Agarose gel electrophoresis showed the intact, sharp and clear
DNA bands with little degradation. The extracted DNA could be used in restriction analysis
and as template in polymerase chain reaction (PCR) using many types of primer including
housekeeping gene, universal, SCAR, EST and SSR primers in both of single and multiplex
PCR. It indicated that the DNA extracted by SDS/NaCl+PVP method had purity, good quality
and was suitable for molecular analyses such as restriction digestion and PCR
amplification.The wholeprocessof DNA extractionusing SDS/NaCl+PVP methodtook about
48 minutes.
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