VEGETOS: Международный журнал исследований растений

Fast Protocol For Extraction of DNA From Arid Adapted Prosopis Leaves Without Liquid Nitrogen

Claudia Yared Michel-López, Daniel González-Mendoza and Onecimo Grimaldo-Juarez

Fast Protocol For Extraction of DNA From Arid Adapted Prosopis Leaves Without Liquid Nitrogen

The extraction of high quality genomic DNA for PCR amplification from Prosopis sp., is complicated due to the presence of a high percentage of secondary metabolites which bind to or co-precipitate with nucleic acids. In the present study we report a modified sodium dodecyl sulfate/phenol protocol that includes elimination of liquid nitrogen in maceration process, β-mercaptoethanol in buffer extraction and ethanol precipitation step. The A260/280 absorbance ratios of isolated DNA were around 2.0 to 1.9, suggesting that the DNA fraction was pure and may be used for further PCR analysis. The DNA isolated by this protocol is of sufficient quality for molecular applications; this technique could be applied to other organisms that have similar substances that hinder DNA extraction. Finally, this proposal represents an alternative fast, cheap, and effective for isolation of genomic DNA from fresh leaves of Prosopis spp., even in low technology laboratories .