Optimization of quality DNA isolation protocol from various mucilage rich cultivated and wild Abelmoschus sp. and its validation through PCR amplifcation

Abstract

Okra is such a genomically less explored crop that, many labs are still trying for the optimization of quality DNA isolation protocol for the efficient use in the contemporary genomic studies. In this study, we have optimized a quick and reproducible DNA isolation protocol for the isolation of quality genomic DNA from different tissues of two cultivated (Abelmoschus esculentus and A. caillei) and seven wild okra species (A. manihot, A. moschatus, A. tetraphyllus, A. tuberculatus, A. ficulneus, A. rugosus and A. angulosus), including related species Hibiscus cannabinus. The quality of isolated DNA was also confirmed using PCR amplification of various DNA markers like Inter Simple Sequence Repeat (ISSR), Random Amplified Polymorphic DNA (RAPD), Start Codon Targeted (SCoT) and Expressed Sequence Tag-Simple Sequence Repeat (EST-SSR). This is probably the first report wherein the DNA isolation protocol has been optimized for nine Abelmoschus species along with a related species Hibiscus. Thus, good quality and quantity of DNA can be isolated in okra, with the care that it should be performed using appropriate plant tissue, either very young leaves or etiolated seedlings, and well optimized DNA isolation protocol as prescribed in this study.