Title : Temperature and Concentration Storage Variation of Sugarcane RNA towards Reverse Transcriptase PCR approach
Abstract :Containing high-quality RNA is the basis of plant molecular biology research, plant genetics and other physiological investigations. At present, several plant RNA isolation methods have been utilized to received high-quality RNA yield. However, there are not much research data regarding RNA resistance after RNA isolation stage. This study attempts to elaborate on the effects of RNA storage on RNA quality for general molecular biology experiments such as Reverse Transcriptase PCR to obtain specific DNA fragments. Temperature and concentration were chosen because they are the most basic factors that can affect RNA quality. The samples were checked using Nanodrop spectrophotometer and gel electrophoresis for testing purity, quantity and integrity of RNA. Based on this experiment, total RNA from sugarcane can be stored at -40 °C and -20 °C for a month without significant quality quantity reduction. Both storage temperatures allow the quality of RNA to be maintained at a sufficient level for use in cDNA synthesis and gene-specific amplification without additional treatment. Storing pure RNA at a sufficiently high concentration would be more advantageous for the next experimental stage