Optimizing Total RNA Extraction Method for Animal Tissues
Abstract
Background. the extraction of high-quality total RNA is pivotal for advanced RNA molecular studies, such as Next-generation sequencing and Microarray DNA hybridization. Despite the development of numerous extraction methods in recent decades, like the CTAB and the Traditional Trizol reagent methods, their complexity and high costs often impede their application in small-scale laboratories. Therefore, a practical and economical method for RNA extraction that maintains high standards of efficiency and quality need to be provided.Method. this study proposes enhancements to the Traditional Trizol method through the incorporation of guanidine isothiocyanate (GITC-T method) and/or sodium dodecyl sulfate (SDS-T method). The objective is to optimize RNA extraction from animal tissues. We evaluated the effectiveness of these modified methods in comparison to the Traditional Trizol method using electrophoresis, Q-PCR, and RNA-Seq.Result. electrophoresis and RNA-Seq demonstrated that the GITC-T method yielded RNA with higher integrity and purity while the consistency in RNA quality between the two methods was confirmed . Furthermore, Q-PCR indicated that the GITC-T method achieved higher yields and greater transcript abundance of total RNA from the same types of animal samples than the Trizol methodConclusion. the GITC-T method not only yields higher purity and quantity of RNA but also reduces reagent consumption and overall costs, thereby presenting a more feasible option for small-scale laboratory settings.