Optimization of the genomic DNA extraction in some mosses

The presence of organic compounds and high amount of secondary metabolites (polysaccharides, phenolic component, etc.) in mosses cause difficulties in DNA extraction that are followed by problems in PCR reactions. In lower plants, various methods have been used for DNA extraction including silica gel and different commercial kits. These methods mostly use hazardous (like phenol or liquid nitrogen) or costly (proteinase K) materials. Commercial kits are high cost. In order to develop an appropriate and cost effective procedure for DNA extraction in lower plants, the CTAB protocol was modified. Triton X-۱۰۰, SDS, activated charcoal and ammonium acetate were used for the elution of the contaminations instead of the hazardous and risky materials. The method was compared with three extraction kits (Vivantus, Biobasic, and Rana), and tested on nine species of mosses including Neckera complanata, Anomodon viticulosus,Trichostomum brachydontium,Dicranum scoparium, Tortula sp., Plagiomnium cuspidatum,Homalothecium sericeum,Eurhynchium sp., and Neckera crispa from Iran. The quality and quantity of the extracted DNA was examined with spectrophotometer and agarose gel electrophoresis. The lack of expensive proteinase K in this procedure had no unfavorable effect on the final results and helped to decrease the costs.

Keywords:

Expensive , non-flowering plants , proteinase K , risky materials , Triton X-۱۰۰