Abstract
Different protocols are usually used for extracting total deoxyribonucleic acid (DNA) from different plant species of same order and DNA of the associated viruses. Here, we describe a rapid, efficient and universal protocol for isolating total DNA from the members of Zingiberales which harbor a high amount of polysaccharides and secondary metabolites. DNA isolated with this protocol was successfully used for PCR based downstream applications viz. random amplified polymorphic DNA (RAPD), Inter-simple sequence repeats (ISSR), DNA barcoding gene (Internal transcribed spacer and trnl-f) amplification and detection of the viruses.
Highlights
Molecular techniques require isolation of genomic deoxyribonucleic acid (DNA) of suitable purity
The isolated high quality genomic DNA is amenable to random amplified polymorphic DNA (RAPD) (Random amplified Polymorphic DNA), ISSR (Inter-simple sequence repeats), amplification of plant barcode genes (ITS and trnL-F) and detection of virus associated with Musa with reduced cost and health concerns
The average yield of total nucleic acid from 1 g of plant material using our method ranged from 879.59 μg/μl to 2350.81 μg/μl which is much higher than those obtained with kits and normal cetyl trimethylammonium bromide (CTAB) method
Summary
Molecular techniques require isolation of genomic DNA of suitable purity. The isolation of good quality DNA is the prerequisite for molecular research. The cetyl trimethylammonium bromide (CTAB) method and its modifications have been used to obtain good quality total DNA for polymerase chain reaction (PCR) based downstream applications.
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