Assessment of Small Scale Methods for Quick Extraction of Genomic DNA from Leaf Tissue of Sugarcane

Abstract

Molecular studies to characterize genetic resources of sugarcane for use in breeding programmes require DNA extraction from several clones simultaneously so the DNA isolation procedures should be rapid, cheap, simple and easy to perform. Seven rapid protocols differing with respect to the composition of extraction buffers and DNA precipitation methods were compared to assess the quantity and quality of genomic DNA extracted from leaf material of different sugarcane genotypes. The DNA obtained was quantified and tested for quality, restriction digestion and PCR amplification using RAPD, SSR and ISSR primers. Two CTAB based methods essentially standardized over sugarcane were found most suitable for both large and small-scale isolation of DNA based on the purity, quality, quantity, applicability for PCR and stability upon storage. A third protocol also produced DNA of sufficient utility, however it requested further modifications.