Two mini-preparation protocols to DNA extraction from plants with high polysaccharide and secondary metabolites

Abstract

Were standardized two previously reported standard plant DNA extraction methods, but improved them on mini preparations to use the samples for population genetic analysis. The combination of CTAB lysis procedure-solvent extraction and DNA column purification (DNeasy plant mini kit modification) enables a faster and reliable DNA extraction from all samples tested:

Piper, Quercus, Zea and cacti species (considered “hard” extraction species), and this protocol uses smaller tissue samples than other mini or midi-prep protocols. We obtained high quality and DNA yields in all samples tested. This alternative protocol (CTAB lysis-solvent extraction based) is an excellent option if there are many samples to process and it is also a non expensive protocol. This method also produces good DNA quality but fewer yields. Both two protocols produce reproducible PCR pattern-bands amplification with all the genetic markers tested (RAPD’s and microsatellites). The DNA obtained was used in other molecular biology standard analysis methods, like enzymptic restriction patterns, ligation, sequencing and cloning with good results too.