Graphene Oxide in Modified LiCl Precipitation Procedure as an Efficient Method to Extract RNA with High Resolution from Zelkova carpinifolia Tissues

Abstract

The objective of present study was to determine the appropriate procedure to extract RNA with high resolution from Zelkova carpinifolia tissues which is known as one of near to threatened forest trees. The total RNA isolation was performed through commercially developed kits and modified LiCl precipitation method in callus, stem and leaves of Z. carpinifolia. Moreover, modified LiCl precipitation method was used as appropriate method with Graphene oxide. It was found that the modified LiCl precipitation method could result high efficiency compared to other used commercial kits. The results showed that the isolated RNA from callus was 271.2 (by modified LiCl precipitation) > 12.12 (Biozol kit) > 9.6 (RNeasy kit) > 5.16 µg/g (Biozol with PVP and β-ME). Mostly, the same trend was occurred for stem and leaf tissues, as well. The high performance of modified LiCl method was confirmed by electrophoresis technique through the clear bands compared to other mentioned methods. Also, high efficiency of used methodology was confirmed by cDNA synthesis, RT-PCR and quantitative real-time PCR (QRT-PCR). Graphene oxide with modified LiCl precipitation revealed better and sharp bands. Finally, an efficient method with Graphene oxide was optimized for high quality and quantity RNA extraction from woody plants.