(1) Present address: Department of Trait Genetics and Technologies, Dow AgroSciences LLC, Building 306/C2, 9330 Zionsville Road, 46268 Indianapolis, IN
Abstract:
The extraction of high-quality genomic DNA for PCR amplification from sunflower (Helianthus annuus) and cotton (Gossypium spp.) is challenging because of the presence of polysaccharides, secondary metabolites, and polyphenolics in the tissues.
A high-throughput DNA extraction protocol was needed in our laboratory for simple sequence repeats (SSR)-marker screening
and other molecular analyses that do not require organic extraction steps of phenol or chloroform. Here we describe 2 improved
highthroughput protocols for DNA extraction and in-PCR modification that result in successful PCR amplification of sunflower
and cotton. While the sunflower DNA extraction protocol uses reducing agents such as sodium metabisulfite and dithiothreitol
(DTT), the cotton protocol uses polyvinylpyrrolidone (PVP) in PCR reactions and reducing agents in the DNA extraction procedure.