Not Available: Isolation of high quality, intact high molecular weight genomic DNA from plants which
are rich in polysaccharides, polyphenols, secondary metabolites and chemical heterogeneity is an
immense problem in the field of plant biology. Several protocols have been developed for
eliminating these tricky elements during the extraction of DNA, but none is found to be
universally applicable. The purpose of the present study was to develop a reliable protocol for
extracting high quality genomic DNA from polyphenols, polysaccharides and secondary
metabolites rich plants like sweet sorghum. We made seven critical modifications to the available
CTAB method to isolate genomic DNA from 25- and 60-d-old transgenic sweet sorghum leaf
tissues. The yield of DNA ranged from 9.2– 10.2 µg from 200 mg of leaf tissue. An absorbance
value of 1.8 at A260/A280 indicates that it’s free from RNA and protein contamination. PCR analysis
using bar primers shows a consistent and reliable amplification product at 475 bp. This method is
highly suitable for extracting high quality genomic DNA from plants with high levels of
polysaccharides and polyphenolics without blending commercial kits. Our protocol facilitates the
processing of large number of plant samples for genomic analysis, mapping and next generation
sequencing.Not Availabl
