An improved method for DNA isolation from polyphenols rich leaf samples of Annona squamosa L.

Abstract

A reliable and efficient method for isolating Annona squamosa L. genomic DNA, free from polyphenols and polysaccharides has been developed. Different methods involving use of CTAB and SDS with or without modifications were used. A CTAB based extraction method which uses diatomite to remove polyphenols and polysaccharides proved to be the best. This method allowed recovery of good quality DNA in sufficient quantity suitable for complete digestion by restriction endonucleases and amplifiable in polymerase chain reaction as compared to other methods.

Molecular characterization of diversity in Annona squamosa L.: A high value tropical medicinal plant

“…Conservation programs are already using data generated by molecular techniques to evaluate the genetic structure and diversity of natural populations. 4,33 In present study 35 primers (19RAPD +15ISSR) primers produced 73.9% polymorphism. Similar results with 73% polymorphism were reported by Bharad et al 35 by analyzing 11 genotypes of A. squamosa using five RAPD primers.…”

“…Genomic DNA was extracted from this new improved method using cetyltrimethylammonium bromide (CTAB) with purification based on diatomite. 33 Optimized PCR Conditions Optimized Conditions for RAPD Analysis Of the 53 RAPD primers initially screened, 19 produced clear and scorable amplification products (Figure 4). These primers yielded a total of 148 amplicons with an average of 8 bands per primer.…”