Fast and Easy Protocol of DNA Extraction from Fresh and Hard/Dried Plant Samples of Indian Spices
Sudeep Nagaraj *
Sapthagiri Institute of Medical Sciences and Research Centre, Bengaluru, Karnataka, India
Ragunandana Girenahalli
Sridevi Post Graduate Centre, Tumakur, Karnataka, India
Praveen Kumar Kondenahalli Subbarayappa
Sridevi Post Graduate Centre, Tumakur, Karnataka, India
Jagadish Tavarekere Venkataravanappa
SDUAHER, Tamaka, Kolar, Karnataka, India
*Author to whom correspondence should be addressed.
Abstract
Spices identified using basic techniques like magnifying glasses or microscopic instruments. The DNA based spice detection techniques used in food testing labs for the disclosure of adulterants. The molecular techniques like PCR, Sequencing, and HRM etc. enables the exact disclosure of spices. Plants synthesize more amounts of polysaccharides, flavonoids, polyphenols and other secondary metabolites which interfere with the pure and high-quality genomic DNA extraction. Although plant DNA isolation protocols, extracting DNA from dried plant samples is a challenging task. This present research describes an expeditious, reliable SDS buffer protocol suited for extracting DNA from both fresh and dried plants samples rich in polysaccharides and secondary metabolites, and the protocol does not need toxic chemicals like phenols and use of liquid nitrogen. Extracted DNA showed the excellent purity evidenced by UV spectrometer at the ratio of A260/A280 ranging from approximately 1.8-1.85 and the A260/A230 ratio was approximately 2, which explains the preparations are sufficiently free from proteins and polysaccharides. The developed method is satisfactory for both dry and fresh spice samples. The success of the present study is getting high-quality genomic DNA and helps to carry out DNA based experiments with a shorter duration of time.
Keywords: SDS, DNA isolation, corrosive chemicals, dry and fresh plant samples