Optimization of DNA Extraction for ITS/U4U3 analysis of Rhipsalis baccifera

Abstract

Genetic analysis of plants relies on high yields of pure DNA. For Rhipsalis baccifera this represents a great challenge since its plant tissue can accumulate large amounts of mucilage, polysaccharides, polyphenols and secondary metabolites, which co-purify with amplifiable DNA. These contaminating compounds lead to a poor yield and prevent acces to PCR-based analysis. These contaminating compounds lead to a poor DNA yield and prevent access to PCR-based analysis. A number of factors, including choice of plant tissue, tissue preparation, and modifications of the extraction buffer, can impact on DNA extraction process. In this study, four different DNA extraction procedures were tested aiming to develop a simple protocol based on CTAB buffer. The obtained results showed that the use of the outer cuticle of old lyophilized tissue allowed reliable results in R. baccifera plants with a good purity ranging from 1.6 to 1.8 and high DNA yield yield ? 500 ng ?L-1.